EP3416651A1 - Composés stéroïdes c-20, compositions et utilisation de ceux-ci pour traiter un traumatisme craniocérébral (tcc), y compris des commotions cérébrales - Google Patents

Composés stéroïdes c-20, compositions et utilisation de ceux-ci pour traiter un traumatisme craniocérébral (tcc), y compris des commotions cérébrales

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Publication number
EP3416651A1
EP3416651A1 EP16815158.7A EP16815158A EP3416651A1 EP 3416651 A1 EP3416651 A1 EP 3416651A1 EP 16815158 A EP16815158 A EP 16815158A EP 3416651 A1 EP3416651 A1 EP 3416651A1
Authority
EP
European Patent Office
Prior art keywords
alkyl
compound
tbi
crystal
double bond
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP16815158.7A
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German (de)
English (en)
Inventor
Xia LU
Yanfeng Zhang
Daniel E. Levy
Michael LEWANDOWSKI
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Prevacus Inc
Original Assignee
Prevacus Inc
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Publication date
Application filed by Prevacus Inc filed Critical Prevacus Inc
Publication of EP3416651A1 publication Critical patent/EP3416651A1/fr
Withdrawn legal-status Critical Current

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
    • A61K31/57—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of two carbon atoms, e.g. pregnane or progesterone
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J1/00—Normal steroids containing carbon, hydrogen, halogen or oxygen, not substituted in position 17 beta by a carbon atom, e.g. estrane, androstane
    • C07J1/0051—Estrane derivatives
    • C07J1/0062—Estrane derivatives substituted in position 17 alfa not substituted in position 17 beta
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J1/00—Normal steroids containing carbon, hydrogen, halogen or oxygen, not substituted in position 17 beta by a carbon atom, e.g. estrane, androstane
    • C07J1/0051—Estrane derivatives
    • C07J1/0081—Substituted in position 17 alfa and 17 beta
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J13/00—Normal steroids containing carbon, hydrogen, halogen or oxygen having a carbon-to-carbon double bond from or to position 17
    • C07J13/007—Normal steroids containing carbon, hydrogen, halogen or oxygen having a carbon-to-carbon double bond from or to position 17 with double bond in position 17 (20)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J15/00—Stereochemically pure steroids containing carbon, hydrogen, halogen or oxygen having a partially or totally inverted skeleton, e.g. retrosteroids, L-isomers
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J21/00—Normal steroids containing carbon, hydrogen, halogen or oxygen having an oxygen-containing hetero ring spiro-condensed with the cyclopenta(a)hydrophenanthrene skeleton
    • C07J21/005—Ketals
    • C07J21/006—Ketals at position 3

Definitions

  • the present invention relates to novel C-20 steroid compounds, compositions and uses thereof for treating, minimizing and/or preventing traumatic brain injury (TBI), including severe TBI, moderate TBI, and mild TBI, including concussions.
  • TBI traumatic brain injury
  • the present invention further relates to polymorphs of ent-19-norprogesterone.
  • TBI traumatic brain injury
  • TBI amongst U.S. military personnel is also a critically important health concern especially for veterans in the Operation Iraqi Freedom (01F) and Operation Enduring Freedom (OEF).
  • DOBIC Defense and Veterans Brain Injury Center
  • DoD Department of Defense
  • 33,149 U.S. military personnel were diagnosed with a TBI in 2011 alone.
  • This number included service members (SMs) in the Army, Navy, Marine Corps, Air Force, and from the active duty and reserve components of the National Guard. See U.S. Dept. of Defense:
  • TBI is a nondegenerative, noncongenital insult to the brain that can result from a bump, blow or jolt to the head or a penetrating head injury that disrupts the normal function of the brain possibly leading to permanent or temporary impairment of cognitive, physical, and psychosocial functions, with an associated diminished or altered state of consciousness. Not all blows or jolts to the head can cause a TBI.
  • the severity of a TBI can range from“mild” to“severe”.
  • A“mild TBI” is characterized as a brief change in mental status or consciousness, whereas a“severe TBI” is characterized as an extended period of unconsciousness or memory loss after the injury.
  • the Glasgow Coma Scale defines the severity of a TBI within 48 hours of injury.
  • moderate to severe brain injuries are defined as follows:
  • Moderate brain injury is defined as a brain injury resulting in a loss of consciousness from 20 minutes to 6 hours and a Glasgow Coma Scale of 9 to 12. See
  • Severe brain injury is defined as a brain injury resulting in a loss of consciousness of
  • Mild traumatic brain injury is defined as the result of the forceful motion of the head or impact causing a brief change in mental status (confusion, disorientation or loss of memory) or loss of consciousness for less than 30 minutes. While MRI and CAT scans are often normal, a person with a mild TBI may remain conscious or may experience a loss of consciousness for a few seconds or minutes. Other symptoms of mild TBI include headache, confusion, difficulty thinking, lightheadedness, dizziness, blurred vision or tired eyes, ringing in the ears, bad taste in the mouth, fatigue or lethargy, frustration, a change in sleep patterns, behavioral or mood swings, memory problems, concentration, attention, or thinking. See http://www.traumaticbraininjury.com/symptoms-of-tbi/mild-tbi- symptoms/.
  • a person with a moderate or severe TBI may present these same symptoms, but may also present a headache that gets worse or does not go away, repeated vomiting or nausea, convulsions or seizures, an inability to awaken from sleep, dilation of one or both pupils of the eyes, slurred speech, weakness or numbness in the extremities, loss of coordination, and increased confusion, restlessness, or agitation. See http://www.ninds.nih.gov/disorders/tbi/tbi.htm
  • a concussion is a type of traumatic brain injury (TBI) caused by a bump, blow or jolt to the head with a temporary loss of brain function. Concussions can also occur from a fall or a blow to the body that causes the head and brain to rattle or move quickly back and forth. See http://www.cdc.goviconcussion/pdf/Fact_Sheet_ConcussTBI-a.pdf. See also Facts about Concussion and Brain Injury at http://www.cdc.goviconcussion/pdf/Fact_Sheet_ConcussTBI- a.pdf.
  • Concussions are defined as a traumatically induced transient disturbance of brain function and involves a complex pathophysiological process and are a subset of MTBI, which are generally self-limited and at the less-severe end of the brain injury spectrum. See Harmon KG et al.:
  • Traumatic_Brain_Injury_and_Persistent_Symptoms Although the term“concussion” is still used in sports literature as interchangeable with“MHI” or“MTBI”, the general clinical medical literature now uses“MTBI” instead. See Barth JT, Varney NR, Ruchinskas RA, Francis JP: Mild head injury: The new frontier in sports medicine. In Varney NR, Roberts RJ. The Evaluation and Treatment of Mild Traumatic Brain Injury. Hillsdale, New Jersey: Lawrence Erlbaum
  • progesterone is a C-21 steroid hormone.
  • the chemical structure for progesterone is as follows:
  • Progesterone is a progestogen, and it is one of the major naturally occurring human progestogens. Progesterone is involved in the female menstrual cycle, pregnancy and
  • Progesterone is naturally produced by the ovaries of mammals, but can also be produced by some plants and yeast.
  • 19-Norprogesterone is a C-20 steroid hormone.
  • the chemical structure for 19- Norprogesterone is as follows:
  • 19-Norprogesterone is believed to be a potent progesten with mineralcorticoid properties and high affinity for the progesterone receptor. See Paris J, Botella J, Fournau P, Bonnet P, Thevenot R: Extinction of mineralocorticoid effects in 19-norprociesterone derivatives:
  • 19-Norprogesterone is a member of the family of 19-nor-corticosteroids that is produced in extra-adrenal tissue in biologically relevant quantities. Levels of this class of steroids are known to be increased and possibly pathogenic in certain states of human hypertension. See Melby JC, Dale SL, Holbrook M, Griffing GT: 19-Nor-corticosteroids in experimental and human hypertension. Clin Exp Hypertens A; 4 (9 ⁇ 10):1851-67 (1982).
  • progesterone and its analogues have many medical applications, both to address acute situations and to address the long-term decline of natural progesterone levels. Other uses of progesterone include, for example, the prevention of preterm birth, to control anovulatury bleeding, to increase skin elasticity and bone strength, and to treat multiple sclerosis.
  • progesterone may be useful for the treatment of traumatic brain injury (TBI), which may result in substantial and sustained improvements in cytologic, morphologic, and functional outcomes. See Schumacher M, Weill-Engerer S, Liere P, et al.:
  • progesterone may reduce poor outcomes following traumatic brain injury by inhibiting inflammatory factors (TNF-a and IL-13) and subsequently reducing brain edema. See Pan, D., et al.: Biomed Environ Sci.
  • progesterone may effectively attenuate edema in both rodent sexes following injury (Djebaili, M., et al.: J Neurotrauma. 22, 106-118 (2005).
  • progesterone and its metabolites has focused on the treatment of TBI...
  • This line of research originated when researchers...found that, after bilateral contusion injury to the medial frontal cortex in young adult male and female rats, 5 days of treatment with progesterone significantly improved spatial learning and sensory performance, compared with controls given injections of the vehicle alone.
  • the first successful clinical trial for the treatment of TBI in more than 30 years of research was recently completed.
  • the SyNAPSe® study s Independent Data and Safety Monitoring Board (DSMB) has released six analyses of the trial’s safety data over the course of the study, concluding each time that SyNAPSe® should continue to its intended completion.
  • the DSMB s formal interim analysis of primary six-month efficacy data from 400 SyNAPSe patients, conducted in January 2013, concluded that there was no reason to stop the study for futility.[and]
  • the SyNAPSe® study is endorsed by the American Brain Injury Consortium (ABIC) and the European Brain Injury Consortium (EBIC).” See
  • 19-norprogesterone and its analogs may have medical applications.
  • this class of compounds is believed to facilitate axon remylination. See Hussain R, EI-Etr M, Gaci 0, Rakotomamonjy J, Macklin WB, Kumar N, Sitruk-Ware R, Schumacher M, Ghoumari AM: “Progesterone and Nestorone facilitate axon remyelination: a role for progesterone receptors”, Endocrinology, 152 (10): 3820-31 (2011). Additionally, this class of compounds has been studied as potential oral contraceptives.
  • Progesterone exists in a non-naturally occurring enantiomeric form known as ent- progesterone:
  • ent-Progesterone is believed to have equal efficacy to natural progesterone in reducing cell death, brain swelling, and inflammation while the enantiomer has three times the antioxidant activity of racemate under certain conditions.
  • ent-Progesterone is believed to have fewer sexual side effects such as suppression of spermatogenesis; inhibition of the conversion of testosterone to dihydrotestosterone; reduction in the size of the testes, epididymis, and leydig cells; and no hyper-coagulative risk as may be seen with natural progesterone.
  • utilities for ent-progesterone have been described in U.S. Patent Application No. 13/645,881, which was filed on October 5, 2012 and is entitled“Nasal Delivery Mechanism for Prophylatic and Post-Acute Use for Progesterone and/or Its Enantiomer for Use in Treatment of Mild
  • Traumatic Brain Injuries U.S. Patent Application No. 13/645,854, which was filed on October 12, 2012 and is entitled“Prophylactic and Post-Acute Use of Progesterone and Its Enantiomer to Better Outcomes Associated with Concussion,” and U.S. Patent Application No. 13/645,925, which was filed on October 12, 2012 and is entitled“Prophylactic and Post- 15 Acute Use of Progesterone in Conjunction with Its Enantiomer for Use in Treatment of Traumatic Brain Injuries, the entire contents and disclosures each of which are incorporated herein by reference in their entireties. See also VanLandingham et al.: The enantiomer of progesterone acts as a molecular neuroprotectant after traumatic brain injury, Neuropharmacology, 51:1078-1085 (2006).
  • progesterone is not suitable for long-term chronic administration for indications outside of hormone replacement therapy and contraception.
  • TBI moderate to severe traumatic brain injury
  • MTBI in the U.S. population including among those who served in the military, is a public health problem, the magnitude and impact of which are underestimated by current civilian and military surveillance systems.
  • TBI traumatic brain injury
  • MTBI in the U.S. population, including among those who served in the military, is a public health problem, the magnitude and impact of which are underestimated by current civilian and military surveillance systems.
  • TBI traumatic brain injury
  • concussions to identify preventable and modifiable risk factors
  • develop and test strategies to reduce MTBIs in civilian and military life develop and test strategies to reduce MTBIs in civilian and military life, and improve health and social outcomes and quality of life for those who sustain these injuries.
  • novel MTBI treatments that are effective, that can be conveniently administered on demand, that are tissue-specific and/or that do not induce side effects, such as those commonly associated with progester
  • the present invention overcomes many of the disadvantages and shortcomings associated with the current state of mild traumatic brain injury (MTBI) treatment through the discovery of certain novel C-20 steroid compounds, namely, ent-19-norprogesterone (PRV-002), compositions and methods of use that are believed to be effective in the treatment of MTBI, including concussions a subset thereof, that can be administered either in accordance with a prescribed treatment regimen or conveniently on demand.
  • the C-20 steroid compounds and/or compositions thereof of the present invention are believed to be tissue-specific and/or do not induce side effects, such as those associated with progesterone or the reproductive system.
  • the C-20 steroid compounds and/or compositions thereof of the present invention can be conveniently administered by any route of administration, especially topically, e.g., pernasally, buccally and/or sublingually, on demand to deliver an effective amount to effectively and/or prophylactically treat and or prevent MTBI.
  • the C-20 steroid compounds and compositions thereof as contemplated by the present invention are believed to be tissue-specific in the brain for treating MTBI and/or do not induce side effects commonly associated with progesterone or the reproductive system.
  • C-20 steroid compounds of the present invention have a common chemical structure as shown by Formula I below:
  • X is O, N or S
  • Y is O, N or S; or, YR 8 R 10 is absent;
  • R 1 , R 2 , R 5 , and R 6 are independently H, C 1 -C 6 alkyl, halogen, OR 12 , NR 13 R 14 , SR 15 ,
  • R 4 is H or C 1 -C 6 alkyl; R 4 together with R 3 and X forms an optionally substituted 5-6 membered heterocycle containing 1-2 nitrogen, oxygen or sulfur atoms; or
  • R 3 is H or C 1 -C 6 alkyl; R 3 together with R 4 and X forms an optionally substituted 5-6 membered heterocycle containing 1-2 nitrogen, oxygen or sulfur atoms, or R 3 is absent;
  • R 7 is absent, H, C(O)-C 1 -C 6 alkyl, C 1 -C 6 alkyl; or R 7 and R 4 together form a double bond;
  • R 8 is H, C(O)-C 1 -C 6 alkyl, C 1 -C 6 alkyl; R 8 together with R 9 and Y forms an optionally substituted 5-6 membered heterocycle containing 1-2 nitrogen, oxygen or sulfur atoms, or R 8 is absent;
  • R 9 is H or C 1 -C 6 alkyl; R 9 together with R 8 and Y forms an optionally substituted 5-6 membered heterocycle containing 1-2 nitrogen, oxygen or sulfur atoms; R 9 and R 11 together form a double bond;
  • R 10 is absent, H, C(O)-C 1 -C 6 alkyl, C 1 -C 6 alkyl; or R 10 and R 11 together form a double bond;
  • R 11 is H or C 1 -C 6 alkyl; or R 11 and R 10 together form a double bond;
  • R 11 and R 9 together form a double bond;
  • R 12 , R 13 , R 14 , R 15 , R 16 and R 17 are independently H, C(O)-C 1 -C 6 alkyl or C 1 -C 6 alkyl; and the dotted line indicates the presence of either a single or a double bond wherein the valences of a single bond are completed by hydrogens.
  • the C-20 steroid compounds of the present invention as depicted in Formula I possess the stereochemical configurations of natural steroids.
  • the C-20 steroid compounds of the present invention, as shown in Formula I may be racemic.
  • the C-20 steroid compounds of the present invention, as illustrated by Formula I may have stereochemical configurations that are opposite to that of natural steroids.
  • Ent-19-norprogesterone has a molecular formula of C 20 H 28 O 2 and a molar mass of 300.435 g/mol.
  • the chemical names for ent-19-norprogesterone include ent-19-norpregn-4-ene-3,20-dione.
  • the chemical structure of ent-norprogesterone is as follows:
  • ent-19- norprogesterone exist in at least two polymorphic forms, polymorph A and polymorph B, each having distinctively different physical properties, and methods of making same.
  • the ent-19- norprogesterone polymorphs are useful in the treatment of TBI, including severe TBI, moderate TBI and mild TBI, including concussions.
  • the present invention contemplates ent-19- norprogesterone in crystalline and amorphous forms, and preferably in crystalline form.
  • pure, single polymorphs as well as mixtures comprising two or more different polymorphs are contemplated.
  • a pure, single polymorph may be substantially free from other polymorphs.
  • substantially free means that other polymorph(s) are present in an amount less than about 15 weight percent, more preferably less than about 10 weight percent, even more preferably less than about 5 weight percent, still more preferably less than about 2 weight percent, and most preferably less than about 1 weight percent.
  • someone with ordinary skill in the art would understand the phrase "in an amount less than about 15 weight percent” to mean that the polymorph of interest is present in an amount of more than about 85 weight percent.
  • the phrase “less than about 10 weight percent” would mean that the polymorph of interest is“substantially pure” in an amount of more than about 90 weight percent, and so on and so forth.
  • “substantially pure” means that the polymorph of interest is present in an amount of at least about 85 weight percent, more preferably at least about 90 weight percent, even more preferably at least about 95 weight percent, still more preferably at least about 98 weight percent, even more preferably at least about 98.8 weight percent, and most preferably at least about 99 weight percent.
  • the term“substantially similar” means an analytical spectrum, such as XRD pattern, Raman spectroscopy, microscopic images, particle distribution, and etc., which resembles the reference spectrum to an appreciable degree in both the peak locations and their intensity.
  • the Type A polymorph is substantially free of other ent-19- norprogesterone polymorphs.
  • substantially free means comprising less 15 weight percent, more preferably less than about 10 weight percent, even more preferably less than about 5 weight percent, still more preferably less than about 2 weight percent, and most preferably less than about 1 weight percent ent-19-norprogesterone polymorphs.
  • the Type A polymorph has a purity of greater than or equal to at least about 90 weight percent, even more preferably at least about 95 weight percent, still more preferably at least about 98 weight percent, even more preferably about 98.8 weight percent, and most preferably at least about 99 weight percent.
  • the Type A ent-19-norprogersterone polymorph exhibits about 15 XRPD peaks, or which about 14 XRPD peaks have an intensity (counts) of at least about 2500 at XRPD peak positions of about 11.2 ⁇ 1.0 degrees, 12.8 ⁇ 1.0 degrees, 13.9 ⁇ 1.0 degrees, 16.4 ⁇ 1.0 degrees, 17.3 ⁇ 1.0 degrees, 18.2 ⁇ 1.0 degrees, 21.1 ⁇ 1.0 degrees, 23.9 ⁇ 1.0 degrees, 25.1 ⁇ 1.0 degrees, 26.2 ⁇ 1.0 degrees, 26.6 ⁇ 1.0 degrees, 29.3 ⁇ 1.0 degrees, 33.8 ⁇ 1.0 degrees and 34.8 ⁇ 1.0 degrees 2-theta (deg), and of which 9 XRPD peaks have an intensity (counts) of at least about 5000 at XRPD peak positions of about 11.2 ⁇ 0.5 degrees, 12.8 ⁇ 0.5 degrees, 16.4 ⁇ 0.5 degrees, 17.3 ⁇ 0.5 degrees, 18.2 ⁇ 0.5 degrees, 21.1 ⁇ 0.5 degrees, 26.2 ⁇ 0.5 degrees, 29.3 ⁇ 0.5 degrees, 29
  • ent-19-norprogersterone polymorph Type A exhibits an X-ray powder diffraction pattern which is substantially similar to the X-ray powder diffraction pattern depicted in FIG.12. In still another embodiment, ent-19-norprogersterone polymorph Type A exhibits an X-ray powder diffraction pattern which is substantially similar to the X-ray powder diffraction pattern depicted in FIG.14. In still another embodiment, ent-19-norprogersterone polymorph Type A exhibits an X-ray powder diffraction pattern which is substantially similar to X-ray powder diffraction pattern depicted in FIG.21. In still another embodiment, ent-19- norprogersterone polymorph Type A exhibits an X-ray powder diffraction pattern which is substantially similar to the X-ray powder diffraction pattern depicted in FIG.23.
  • ent-19-norprogersterone polymorph Type A exhibits TGA/DSC curves which is substantially similar to the TGA/DSC curves depicted in FIG.15, a microscopic image which is substantially similar to the microscopic image depicted in FIG.16, a particle size distribution which is substantially similar to the particle size distribution depicted in FIG.17A and /or FIG.17B, and a microscopic image which is substantially similar to the microscopic image depicted in FIG.18A, FIG.18B, FIG.18C and/or FIG.18D.
  • ent-19-norprogersterone polymorph Type A exhibits at least one or more substantially similar XRPD peak positions, preferably any 1 to 4 substantially similar XRPD peak positions, more preferably any 1 to 9 substantially similar XRPD peak positions, and most preferably any 1 to anyone of the 15 substantially similar XRPD peak positions, as depicted on FIG.12 and/or FIG.14, wherein each such substantially similar XRPD peak position is at about ⁇ 0.5 degrees, more preferably ⁇ 0.4 degrees, even more preferably ⁇ 0.3 degrees and most preferably ⁇ 0.2 degrees of its XRPD peak position as depicted in FIG 12 or FIG 14, at 6.2, 9.2, 12.9, 14.0.15.3, 16.6, 17.5, and 18.4 ⁇ 0.2 degrees 2-theta.
  • the present invention provides a crystalline form of ent-19- norprogesterone selected from the group consisting of a crystalline form having an X-ray powder diffraction pattern substantially the same as
  • an X-ray powder diffraction pattern having at least four 2 ⁇ ° peaks selected from the group consisting of about 11.2 ⁇ 1.0 degrees, 12.8 ⁇ 1.0 degrees, 13.9 ⁇ 1.0 degrees, 16.4 ⁇ 1.0 degrees, 17.3 ⁇ 1.0 degrees, 18.2 ⁇ 1.0 degrees, 21.1 ⁇ 1.0 degrees, 23.9 ⁇ 1.0 degrees, 25.1 ⁇ 1.0 degrees, 26.2 ⁇ 1.0 degrees, 26.6 ⁇ 1.0 degrees, 18.8 ⁇ 1.0 degrees, 29.3 ⁇ 1.0 degrees, 33.8 ⁇ 1.0 degrees and 34.8 ⁇ 1.0 degrees 2-theta (deg);
  • an X-ray powder diffraction pattern having at least four 2 ⁇ ° peaks selected from the group consisting of about 11.2 ⁇ 0.2 degrees, 12.8 ⁇ 0.2 degrees, 13.9 ⁇ 0.2 degrees, 16.4 ⁇ 0.2 degrees, 17.3 ⁇ 0.2 degrees, 18.2 ⁇ 0.2 degrees, 18.8 ⁇ 0.2 degrees, 21.1 ⁇ 0.2 degrees, 23.9 ⁇ 0.2 degrees, 25.1 ⁇ 0.2 degrees, 26.2 ⁇ 0.2 degrees, 26.6 ⁇ 0.2 degrees, 18.8 ⁇ 0.2 degrees, 29.2 ⁇ 0.5 degrees, 33.8 ⁇ 0.2 degrees and 34.8 ⁇ 0.2 degrees 2-theta (deg);
  • an X-ray powder diffraction pattern having at least four 2 ⁇ ° peaks selected from the group consisting of about 11 ⁇ 1.0 degrees, about 13 ⁇ 1.0 degrees, about 14 ⁇ 1.0 degrees, about 16.5 ⁇ 1.0 degrees, about 17 ⁇ 1.0 degrees, about 18 ⁇ 1.0 degrees, 19 ⁇ 1.0 degrees, about 21 ⁇ 1.0 degrees, about 24 ⁇ 1.0 degrees, about 25 ⁇ 1.0 degrees, about 26 ⁇ 1.0 degrees, about 26.5 ⁇ 1.0 degrees, about 29 ⁇ 1.0 degrees, about 30 ⁇ 1.0 degrees, about 34 ⁇ 1.0 degrees, about 38 ⁇ 1.0, degrees and about 35 ⁇ 1.0 degrees 2-theta (deg).
  • the Type B ent-19-norprogersterone polymorph exhibits about 6 XRPD peaks which have an intensity (counts) of at least about 250 at XRPD peak positions of about 13 ⁇ 1.0 degrees, 14.5 ⁇ 1.0 degrees, 15.2 ⁇ 1.0 degrees 16.8 ⁇ 1.0 degrees, 18.5 ⁇ 1.0 degrees and 21.8 ⁇ 1.0 degrees, 2-theta (deg), as depicted in FIG.19, or about 6 XRPD peaks which have an intensity (counts) of at least about 250 at XRPD peak positions of about 13 ⁇ 0.5 degrees, 14.5 ⁇ 0.5 degrees, 15.2 ⁇ 0.5 degrees, 16.8 ⁇ 0.5 degrees, 18.5 ⁇ 0.5 degrees and 21.8 ⁇ 0.5 degrees, 2- theta (deg), as depicted in FIG.19.
  • ent-19-norprogersterone polymorph Type B exhibits an X-ray powder diffraction pattern which is substantially similar to the ent-19-norprogersterone polymorph Type B exhibits an X-ray powder diffraction pattern depicted in FIG.12.
  • ent-19-norprogersterone polymorph Type B exhibits an X-ray powder diffraction pattern which is substantially similar to the ent-19-norprogersterone polymorph Type B exhibits an X-ray powder diffraction pattern depicted FIG.19.
  • ent-19-norprogersterone polymorph Type B exhibits an X-ray powder diffraction pattern which is substantially similar to the ent-19-norprogersterone polymorph Type B exhibits an X-ray powder diffraction pattern depicted FIG.21.
  • ent-19-norprogersterone polymorph Type B exhibits TGA/DSC curves which are substantially similar to the TGA/DSC curves depicted in FIG.20.
  • the present invention provides a crystalline form of ent-19- norprogesterone selected from the group consisting of a crystalline form having an X-ray powder diffraction pattern substantially the same as
  • an X-ray powder diffraction pattern having at least four 2 ⁇ ° peaks selected from the group consisting of about 13 ⁇ 0.2 degrees, 14.5 ⁇ 0.2 degrees, 15.2 ⁇ 0.2 degrees 16.8 ⁇ 0.2 degrees, 18.5 ⁇ 0.2 degrees and 21.8 ⁇ 0.2 degrees, 2-theta (deg).
  • amorphous ent-19-norprogesterone exhibits an amorphous pattern substantially similar to the amorphous pattern depicted in FIG.22. In another aspect of the present invention, amorphous ent-19-norprogesterone exhibits an amorphous pattern substantially similar to the amorphous pattern depicted in FIG.22.
  • the C-20 steroid compounds of Formula I are believed to be useful for treating, minimizing and/or preventing neuronal damage, such as neuronal damage, resulting from various injuries involving TBI, whether the TBI is mild, moderate or severe.
  • An especially preferred treatment in accordance with the present invention is treatment of MTBI, including a concussion, with ent-19-progesterone.
  • a C-20 steroid compound of Formula I may be administered as a single therapeutic agent.
  • the C-20 steroid compounds of Formula I can be administered through routes of administration that include, e.g., oral, sublingual, intravenous, intraperitoneal, subcutaneous, intramuscular, ocular, otic, intranasal, topical, transdermal and rectal routes of administration.
  • routes of administration include, e.g., oral, sublingual, intravenous, intraperitoneal, subcutaneous, intramuscular, ocular, otic, intranasal, topical, transdermal and rectal routes of administration.
  • routes of administration include, e.g., oral, sublingual, intravenous, intraperitoneal, subcutaneous, intramuscular, ocular, otic, intranasal, topical, transdermal and rectal routes of administration.
  • the C-20 compounds of Formula I can be formulated into a novel composition or admixture and administered in the form of, e.g., a tablet, capsule, gelcap, caplet, powder, granule, liquid, solution, suspension,
  • the present invention envisions that the C-20 steroid compounds of Formula I, including ent-19-norprogesterone, may be administered as a formulation that is swallowed, injected, infused, inhaled, applied transdermally or topically, such as applied to the skin, eye, ear, nose, mucosal membrane or any other membrane or inserted into the rectum.
  • administration to treat TBI, especially MTBI, as contemplated by the present invention is the pernasal, inhalation or injection routes of administration.
  • FIG.1 is drawn to a table showing concussion facts
  • FIG.2 is drawn to a drawing illustrating a Morris thigmotaxis water maze
  • FIG.3 is drawn to a chart that shows no significant differences in motor function, as measured by neuroscore, which were observed at 24h post-injury;
  • FIG.4 is drawn to a chart that shows that when rats are treated with either PRV-002 4mg/kg or PRV-00216mg/kg, they have significantly better motor function, compared to vehicle- treated rats, at 48h post-injury. * indicates a significant difference from vehicle-treated, injured rats, p ⁇ 0.05.
  • FIG.5A is drawn to a chart that shows that when treatment is with either PRV-002 4mg/kg or PRV-00216mg/kg, significantly attenuated TBI-related cognitive deficits are observed during trial 1 of the Morris water maze task at 48h post-injury. * indicates a significant difference from vehicle-treated, injured rats, p ⁇ 0.05;
  • FIG.5B is drawn to a chart that shows that when treatment is with either PRV-002 4mg/kg or PRV-00216mg/kg, significantly attenuated TBI-related cognitive deficits are observed during trial 2 of the Morris water maze task at 48h post-injury. * indicates a significant difference from vehicle-treated, injured rats, p ⁇ 0.05;
  • FIG.6A is drawn to a chart that shows vehicle-treated rats spend significantly more time in thigmotaxia compared during sham, PRV-0024mg/kg-treated, or PRV-00216mg/kg-treated rats during trial 1 of the Morris water maze task, 48h post-injury. * indicates a significant difference from vehicle-treated, injured rats, p ⁇ 0.05;
  • FIG.6B is drawn to a chart that shows vehicle-treated rats spend significantly more time in thigmotaxia compared during sham, PRV-0024mg/kg-treated, or PRV-00216mg/kg-treated rats during trial 2 of the Morris water maze task, 48h post-injury. * indicates a significant difference from vehicle-treated, injured rats, p ⁇ 0.05;
  • FIG.7A is drawn to a photograph that shows the nasal mucosa of a rat free of Evans Blue Dye
  • FIG.7B is drawn to a photograph that shows no Evans Blue Dye observable in nasal mucosa of a rat using pipette for IN administration;
  • FIG.7C is drawn to a photograph that shows excellent intranasal penetration observed in nasal mucosa of a rat using micro Atomizer.
  • FIG.8A is drawn to a chart that shows that when injured rats are treated with PRV-002 4mg/kg, they have significantly better cognitive performance, as compared to all other groups, during trial 1 of the Morris water maze task (top). * indicates a significant difference from vehicle-treated, injured rats, p ⁇ 0.05;
  • FIG.8B is drawn to a chart that shows that when injured rats are treated with PRV-002 significant group differences in cognitive performance are not observed during trial 2 of the Morris water maze task (bottom);
  • FIG.9A is drawn to a chart that shows that no significant group differences are observed in time spent in thigmotaxia during trial 1 of the Morris water maze task (top). Uninjured (sham) and PRV-0024mg/kg-treated rats spends significantly less time in thigmotaxia as compared to vehicle-treated injured rats during trial 2 of the Morris water maze task (bottom). * indicates a significant difference from vehicle-treated, injured rats, p ⁇ 0.05;
  • FIG.9B is drawn to a chart that shows rats treated with PRV-002005mg/kg spent significantly more time in thigmotaxia, compared to sham rats, during trial 2. * indicates a significant difference from vehicle-treated, injured rats, p ⁇ 0.05;
  • FIG.10 is drawn to a chart that shows that when rats are treated with PRV-0020.1 mg/kg or PRV-0024mg/kg, they have significantly improved motor function, as compared to vehicle- treated rats at 24h post-injury. All PRV-002 treatment groups had motor performance scores that are not significantly different from sham rats. * indicates a significant difference from vehicle- treated, injured rats, p ⁇ 0.05;
  • FIG.11 is drawn to a chart that shows sham rats and that when rats treated with either PRV-0020.05mg/kg, PRV-0020.1 mg/kg, or PRV-0024mg/kg, the treated rats have significantly better motor function, as compared to vehicle-treated rats at 48h post-injury.
  • PRV-002 PRV-0020.05mg/kg, PRV-0020.1 mg/kg, or PRV-0024mg/kg
  • 0.05mg/kg- and PRV-0021 mg/kg-treated rats have significantly worse performance, compared to sham rats at 48h post-injury. * indicates a significant difference from vehicle-treated, injured rats, p ⁇ 0.05;
  • FIG.12 shows an XRPD pattern overlay of Type A and Type B of PRV-002;
  • FIG.13 shows the interconversion of amorphous PRV-002 Type A and Type B
  • FIG.14 shows an XRPD pattern of PRV-002 Type A (807302-25-A);
  • FIG.15 shows TGA/DSC curves of PRV-002 Type A (807302-25-A);
  • FIG.16 is a microscopic image of PRV-002 Type A (807302-25-A);
  • FIG.17A shows particle size distribution of PRV-002 Type A (807302-25-A) without sonication, (b) sonicated using a power of 30W for 30s;
  • FIG.17B shows particle size distribution of PRV-002 Type A (807302-25-A) sonicated using a power of 30W for 30s;
  • FIG.18A shows a microscopic image of a PRV-002 Type A batch collected from screening
  • FIG.18B shows a microscopic image of a different PRV-002 Type A batch collected from screening
  • FIG.18C shows a microscopic image of another different PRV-002 Type A batch collected from screening
  • FIG.18D shows a microscopic image of yet another different PRV-002 Type A batch collected from screening
  • FIG.19 shows an XRPD pattern of PRV-002 Type B (807302-42-A);
  • FIG.20 shows TGA/DSC curves of PRV-002 Type B (807302-42-A);
  • FIG.21 shows an XRPD pattern overlay of PRV-002 Type B before and after storage at ambient conditions
  • FIG.22 shows an XRPD pattern of an amorphous precipitate sample of PRV-002
  • FIG.23 shows an XRPD pattern overlay of amorphous before and after storage at ambient conditions with characteristic peaks of PRV-002 Type A;
  • FIG.24 shows an XRPD pattern overlay of anti-solvent addition experiments (I/IV) from a PRV-002 Type A sample
  • FIG.25 shows an XRPD pattern overlay of anti-solvent addition (II/IV) from a PRV-002 Type A sample
  • FIG.26 shows aXRPD pattern overlay of anti-solvent addition (III/IV) from a PRV-002 Type A sample
  • FIG.27 shows an XRPD pattern overlay of anti-solvent addition (IV/IV) from a PRV-002 Type A sample
  • FIG.28 shows an XRPD pattern overlay of reverse anti-solvent addition (I/II) from a PRV-002 Type A sample
  • FIG.29 shows an XRPD pattern overlay of reverse anti-solvent addition (II/II) from a PRV-002 Type A sample
  • FIG.30 shows an XRPD pattern overlay of slow cooling (I/II) from a PRV-002 Type A sample
  • FIG.31 shows an XRPD pattern overlay of slow cooling (II/II) from a PRV-002 Type A sample
  • FIG.32 shows an XRPD pattern overlay of slurry at RT (I/III) from a PRV-002 Type A sample
  • FIG.33 shows an XRPD pattern overlay of slurry at RT (II/III) from a PRV-002 Type A sample
  • FIG.34 shows an XRPD pattern overlay of slurry at RT (III/III) from a PRV-002 Type A sample
  • FIG.35 shows an XRPD pattern overlay of slurry at 5 oC (I/II) from a PRV-002 Type A sample
  • FIG.36 shows an XRPD pattern overlay of slurry at 5 oC (II/II) from a PRV-002 Type A sample) ;
  • FIG.37 shows an XRPD pattern overlay of solid vapor diffusion (I/II) from a PRV-002 Type A sample
  • FIG.38 shows an XRPD pattern overlay of solid vapor diffusion (II/II) from a PRV-002 Type A sample
  • FIG.39 shows an XRPD pattern overlay of solution vapor diffusion from a PRV-002 Type A sample
  • FIG.40 shows an XRPD pattern overlay of crystallization induced by polymer mixture A from a PRV-002 Type A sample
  • FIG.41 shows an XRPD pattern overlay of crystallization induced by polymer mixture from a PRV-002 Type A sample
  • FIG.42 shows an XRPD pattern overlay of slow evaporation (I/II) from a PRV-002 Type A sample
  • FIG.43 shows an XRPD pattern overlay of slow evaporation (II/II) from a PRV-002 Type A sample
  • FIG.44 shows an XRPD pattern overlay of grinding from a PRV-002 Type A sample
  • FIG.48 depicts PRV-002 (ent-19-norprogesterone) mean concentrations in plasma of dogs dosed 3 times at 4 hour intervals in one day by IN administration in an amount of about 1mL/nostril at each dosing interval for a total PRV-002 (ent-19-norprogesterone) dose of about 46 mg/dog per dosing interval.
  • administering should be understood to mean providing an active agent of the invention to the subject in need of TBI treatment in a form that can be introduced into that subject's body in a therapeutically useful form and therapeutically effective amount.
  • alkyl refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing no unsaturation, having from one to eight carbon atoms, and which is attached to the rest of the molecule by a single bond, such as illustratively, methyl, ethyl, n-propyl 1-methylethyl (isopropyl), n-butyl, n-pentyl, and 1,1-dimethylethyl (tert- butyl).
  • Amorphous refers to a composition comprising a compound that contains no or too little crystalline content of the compound to yield a discernable pattern by XRPD or other diffraction techniques.
  • glassy materials are a type of amorphous material. Amorphous materials do not have a true crystal lattice, and are glassy, technically resembling very viscous non- crystalline liquids. Glasses may better be described as quasi-solid amorphous material. As is known in the art, an amorphous material refers to a quasi-solid. A compound in an amorphous state may be produced by rapidly evaporating solvent from a solution of a compound, or by grinding, pulverizing or otherwise physically pressurizing or abrading the compound while in a crystalline state.
  • “at least one” means“one or more” of the listed elements.
  • Blood sample refers to whole blood taken from a subject, or any fractions of blood including plasma or serum.
  • compositions and methods include the recited elements, but not excluding others.
  • Consisting essentially of when used to define compositions and methods, shall mean excluding other elements of any essential significance to the combination. For example, a composition consisting essentially of the elements as defined herein would not exclude other elements that do not materially affect the basic and novel characteristic(s) of the claimed invention.“Consisting of” shall mean excluding more than trace amount of other ingredients and substantial method steps recited. Embodiments defined by each of these transition terms are within the scope of this invention.
  • Crystalline refers herein to a material that contains a specific compound or a salt of the compound, which may be hydrated and/or solvated, and has sufficient crystalline content to exhibit a discernable diffraction pattern by XRPD or other diffraction techniques. Crystallines can be characterized by their crystalline structure (X-ray diffraction pattern), their thermal properties (as determined by DSC and TGA), stability, solubility, etc. The X-ray diffraction pattern is presented as characteristic 2 ⁇ ° peaks and one skilled in the art can readily identify a crystalline form of a compound or salt based on the characteristic 2 ⁇ ° peaks of an X-ray diffraction pattern of the polymorph.
  • characteristic peaks are those having a relative intensity of 25% or more. In some embodiments, characteristic peaks are those that have a relative intensity of 10% or more. In some embodiments, characteristic peaks are those that have a relative intensity of 5% or more.
  • a crystalline of a compound or a salt may be characterized by properties including one or more of the following as described in details herein, such as (i) its X-ray powder diffraction pattern (XRPD), (ii) its infrared spectrum (IR), (iii) its differential scanning calorimetry (DSC), (iv) its thermogravimetric analysis (TGA), (v) its vapor sorption curve, such as Gravimetric Vapour Sorption (GVS), and/or (vi) crystal structure, such as unit cell structure.
  • XRPD X-ray powder diffraction pattern
  • IR infrared spectrum
  • DSC differential scanning calorimetry
  • TGA thermogravimetric analysis
  • GVS Gravimetric Vapour Sorption
  • crystal structure such as unit cell structure.
  • a crystalline material that is obtained by direct crystallization of a compound dissolved in a solvent or solvent mixture or solution or interconversion of crystals obtained under different crystallization conditions may have crystals that contain the solvent used in the crystallization.
  • Such compositions may be referred to as a crystalline solvate.
  • the solvent is water
  • such compositions may be referred to as a crystalline hydrate.
  • the specific solvent system and physical embodiment in which the crystallization is performed collectively termed as crystallization conditions, may result in the crystalline material having physical and chemical properties that are unique to the crystallization conditions.
  • cycloalkyl denotes a non-aromatic mono or multicyclic ring system of 3 to 12 carbon atoms such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and examples of multicyclic cycloalkyl groups include perhydronapththyl, adamantyl and norbornyl groups bridged cyclic group or spirobicyclic groups e.g., spiro(4,4)non-2-yl.
  • Disease condition refers to a disease state for which the compounds, compositions and methods of the present invention are being used against.
  • effective amount means any amount or dosage strength of a C- 20 steroid compound of the present invention, especially ent-19-norprogesterone, to treat, minimize and/or prevent traumatic brain injury, including severe, moderate and/or mild TBI, including concussions.
  • Effective amount also means any amount or dosage amount considered by the U.S. Food and Drug Administration (FDA) or other governmental agency or tribunal as being effective to treat, minimize and/or prevent traumatic brain injury, including severe, moderate and/or mild TBI, including concussions.
  • FDA U.S. Food and Drug Administration
  • LG refers to any group that leaves in the course of a chemical reaction involving the group and includes but is not limited to halogen, brosylate, mesylate, tosylate, triflate, p-nitrobenzoate, phosphonate groups, for example.
  • Patient refers to mammals and includes humans and non-human mammals.
  • Polymorph or“polymorphic form” refers to a crystalline form of a substance that is distinct from another crystalline form but that shares the same chemical formula.
  • the different polymorphic forms of the same compound can have an impact on one or more physical properties, such as stability, solubility, melting point, bulk density, flow properties, bioavailability, etc.
  • Pseudopolymorph refers to a crystalline form of a hydrate or solvate of a compound. In contrast to polymorphs, pseudopolymorphs are chemically identical except differ in the amount of water or solvent bound in the crystal lattice. Depending on the solvent used during synthesis and/or crystallization some compounds form hydrates (with water) or solvates (with other solvents) in different stoichiometric ratio. Pseudopolymorphs may show different physical properties like habitus, stability, dissolution rate and bioavailability as known for polymorphs. “Subject” refers to any animal, individual or Patient.
  • “Tautomer” refers to alternate forms of a compound that differ in the position of a proton, such as enol-keto and imine-enamine tautomers, or the tautomeric forms of heteroaryl groups containing a ring atom attached to both a ring—NH— moiety and a ring ⁇ N— moiety such as pyrazoles, imidazoles, benzimidazoles, triazoles, and tetrazoles.
  • the proton of a salt of any compound of the present invention may be in different positions of the molecule.
  • a salt of any compound of the present invention, including ent-19-norprogetserone includes any and all structural variations due to the position of the salt proton unless otherwise indicated.
  • the term“preventing” refers to the prophylactic treatment of a patient in need thereof.
  • the prophylactic treatment can be accomplished by providing an appropriate dose of a therapeutic agent to a subject at risk of suffering from an ailment, thereby substantially averting onset of the ailment.
  • Treatment means any treatment of a disease or disorder in a subject, including preventing, ameliorating and/or protecting against the disease or disorder, that is, (i) causing the clinical symptoms not to develop, (ii) inhibiting the disease or disorder, that is, arresting or suppressing the clinical symptoms or the development of clinical symptoms, and/or (iii) relieving the disease or disorder, that is, causing the regression or arrestation of the clinical symptoms.
  • the particular treatment thus will depend on the disease and state of disease to be targeted and the current or future state of medicinal therapies and therapeutic approaches.
  • a treatment may have associated toxicities.
  • “Therapeutically effective amount” refers to that amount of a compound of this invention, whether used alone or in a suitable pharmaceutical composition that is sufficient to effect treatment, as defined herein, when administered to a subject in need of such treatment, preferably without causing treatment limiting toxicity.
  • the therapeutically effective amount will vary depending upon the subject and disease condition being treated, such as, the weight, the heritage and the age of the subject, the condition of the subject, the severity of the disease condition, the particular compound chosen, the dosage form and the dosing regimen to be followed, timing of administration, the manner of administration and the like, all of which may be determined by one of ordinary skill in the art.
  • a therapeutically effective amount can vary, depending on any of a number of factors, including, e.g., the compound and dosage amount and form selected, the route of administration, the treatment regimen, the condition of the subject, the severity of the disease being treated, as well as other factors understood by those in the art.
  • this invention provides salts, polymorphs, and pseudopolymorphs of the compounds of the present invention.
  • the present invention contemplates any and all salt forms, polymorphs and pseudopolymorphs of the compounds, including ent-19-norprogesterone. Except where noted otherwise, capitalized and non-capitalized forms of all terms fall within each meaning. Singular word forms are intended to include plural word forms and are likewise used herein interchangeably where appropriate and fall within each meaning, unless expressly stated otherwise.All parts, percentages, ratios, etc. herein are by weight unless indicated otherwise.
  • the present invention provides for C-20 steroid compounds having a chemical structure of Formula I:
  • X is O, N or S
  • Y is O, N or S; or, YR 8 R 10 is absent;
  • R 1 , R 2 , R 5 , and R 6 are independently H, C 1 -C 6 alkyl, halogen, OR 12 , NR 13 R 14 , SR 15 ,
  • R 4 is H or C 1 -C 6 alkyl; R 4 together with R 3 and X forms an optionally substituted 5-6
  • R 3 is H or C 1 -C 6 alkyl; R 3 together with R 4 and X forms an optionally substituted 5-6
  • R 7 is absent, H, C(O)-C 1 -C 6 alkyl, C 1 -C 6 alkyl; or R 7 and R 4 together form a double bond;
  • R 8 is H, C(O)-C 1 -C 6 alkyl, C 1 -C 6 alkyl; R 8 together with R 9 and Y forms an optionally substituted 5-6 membered heterocycle containing 1-2 nitrogen, oxygen or sulfur atoms, or R 8 is absent;
  • R 9 is H or C 1 -C 6 alkyl; R 9 together with R 8 and Y forms an optionally substituted 5-6 membered heterocycle containing 1-2 nitrogen, oxygen or sulfur atoms; R 9 and R 11 together form a double bond;
  • R 10 is absent, H, C(O)-C 1 -C 6 alkyl, C 1 -C 6 alkyl; or R 10 and R 11 together form a double bond;
  • R 11 is H or C 1 -C 6 alkyl; or R 11 and R 10 together form a double bond;
  • R 11 and R 9 together form a double bond;
  • R 12 , R 13 , R 14 , R 15 , R 16 and R 17 are independently H, C(O)-C 1 -C 6 alkyl or C 1 -C 6 alkyl;
  • the dotted line indicates the presence of either a single or a double bond wherein the valences of a single bond are completed by hydrogens.
  • the C-20 steroid compounds of Formula I possess the stereochemical configuration of natural steroids.
  • the C-20 steroid compounds of Formula I are racemic.
  • the C-20 steroid compounds of formula I possess a stereochemical configuration that is opposite to that of natural steroids.
  • the present invention provides for C-20 steroid compounds having a chemical structure of Formula II:
  • X is O, N or S
  • Y is O, N or S; or, YR 8 R 10 is absent;
  • R 1 , R 2 , R 5 , and R 6 are independently H, C 1 -C 6 alkyl, halogen, OR 12 , NR 13 R 14 , SR 15 ,
  • R 4 is H or C 1 -C 6 alkyl; R 4 together with R 3 and X forms an optionally substituted 5-6
  • R 3 is H or C 1 -C 6 alkyl; R 3 together with R 4 and X forms an optionally substituted 5-6 membered heterocycle containing 1-2 nitrogen, oxygen or sulfur atoms, or R 3 is absent;
  • R 7 is absent, H, C(O)-C 1 -C 6 alkyl, C 1 -C 6 alkyl; or R 7 and R 4 together form a double bond;
  • R 8 is H, C(O)-C 1 -C 6 alkyl, C 1 -C 6 alkyl; R 8 together with R 9 and Y forms an optionally substituted 5-6 membered heterocycle containing 1 -2 nitrogen, oxygen or sulfur atoms, or R 8 is C 1 -C 6 absent;
  • R 9 is H or alkyl; R 9 together with R 8 and Y forms an optionally substituted 5-6 membered
  • R 10 is absent, H, C(O)-C 1 -C 6 alkyl, C 1 -C 6 alkyl; or R 10 and R 11 together form a double bond;
  • R 11 is H or C 1 -C 6 alkyl; or R 11 and R 10 together form a double bond;
  • R 11 and R 9 together form a double bond;
  • R 12 , R 13 , R 14 , R 15 , R 16 and R 17 are independently H, C(O)-C 1 -C 6 alkyl, C 1 -C 6 alkyl; and the dotted line indicates the presence of either a single or a double bond wherein the valences of a single bond are completed by hydrogens.
  • the C-20 steroid compounds of Formula II possess the
  • the C-20 steroid compounds of Formula II are racemic. In still other embodiments, the C-20 steroid compounds of formula II possess a stereochemical configuration that is opposite to that of natural steroids.
  • the present invention provides for C-20 steroid compounds having a chemical structure of Formula III:
  • X is O, N or S
  • Y is O, N or S; or, YR 8 R 10 is absent;
  • R 4 is H or C 1 -C 6 alkyl; R 4 together with R 3 and X forms an optionally substituted 5-6
  • R 3 is H or C 1 -C 6 alkyl; R 3 together with R 4 and X forms an optionally substituted 5-6 membered heterocycle containing 1-2 nitrogen, oxygen or sulfur atoms, or R 3 is absent;
  • R 7 is absent, H, C(O)-C 1 -C 6 alkyl, C 1 -C 6 alkyl; or R 7 and R 4 together form a double bond;
  • R 8 is H, C(O)-C 1 -C 6 alkyl, C 1 -C 6 alkyl; R 8 together with R 9 and Y forms an optionally substituted 5-6 membered heterocycle containing 1-2 nitrogen, oxygen or sulfur atoms, or R 8 is absent; R 9 is H or C 1 -C 6 alkyl; R 9 together with R 8 and Y forms an optionally substituted 5-6 membered heterocycle containing 1-2 nitrogen, oxygen or sulfur atoms; R 9 and R 11 together form a double bond;
  • R 10 is absent, H, C(O)-C 1 -C 6 alkyl, C 1 -C 6 alkyl; or R 10 and R 11 together form a double bond, R 11 is H or C 1 -C 6 alkyl; or R 11 and R 10 together form a double bond; R 11 and R 9 together form a double bond; and
  • the dotted line indicates the presence of either a single or a double bond wherein the valences of a single bond are completed by hydrogens.
  • the C-20 steroid compounds of Formula III possesses the stereochemical configuration of natural steroids. In other embodiments, the C-20 steroid compounds of Formula III are racemic. In still other embodiments, the C-20 steroid compounds of formula III possess a stereochemical configuration that is opposite to that of natural steroids.
  • the present invention provides for C-20 steroid compounds having a chemical structure of Formula IV:
  • X is O, N or S
  • Y is O, N or S; or, YR 8 R 10 is absent;
  • R 4 is H or C 1 -C 6 alkyl; R 4 together with R 3 and X forms an optionally substituted 5-6
  • R 3 is H or C 1 -C 6 alkyl; R 3 together with R 4 and X forms an optionally substituted 5-6 membered heterocycle containing 1-2 nitrogen, oxygen or sulfur atoms, or R 3 is absent;
  • R 7 is absent, H, C(O)-C 1 -C 6 alkyl, C 1 -C 6 alkyl; or R 7 and R 4 together form a double bond;
  • R 8 is absent, H, C(O)-C 1 -C 6 alkyl, C 1 -C 6 alkyl;
  • R 10 is absent, H, C(O)-C 1 -C 6 alkyl, C 1 -C 6 alkyl; or R 10 and R 11 together form a double bond; and R 11 is H or C 1 -C 6 alkyl; or R 11 and R 10 together form a double bond; R 11 and R 9 together form a double bond; and
  • the dotted line indicates the presence of either a single or a double bond wherein the valences of a single bond are completed by hydrogens.
  • the C-20 steroid compounds of Formula IV possesses the stereochemical configuration of natural steroids. In other embodiments, the C-20 steroid compounds of Formula IV are racemic. In still other embodiments, the C-20 steroid compounds of formula IV possess a stereochemical configuration that is opposite to that of natural steroids.
  • the C-20 steroid compound of Formula I is Compound A:
  • the C-20 steroid compound of Formula I is Compound B:
  • the C-20 steroid compound of Formula I is Compound C:
  • the C-20 steroid compound of Formula I is Compound D:
  • the C-20 steroid compound of Formula I is Compound E:
  • the C-20 steroid compound of Formula I is Compound F:
  • the C-20 steroid compound of Formula I is Compound
  • the C-20 steroid compound of Formula I is Compound H:
  • the C-20 steroid compound of Formula I is Compound I:
  • the C-20 steroid compound of Formula I is Compound J:
  • the C-20 steroid compound of Formula I is Compound K:
  • the C-20 steroid compound of Formula I is Compound L:
  • the C-20 steroid compound of Formula I is Compound M:
  • the C-20 steroid compound of Formula I is Compound N:
  • the C-20 steroid compound of Formula I is Compound O:
  • the C-20 steroid com ound of Formula I is Compound P:
  • the C-20 steroid compound of Formula I is Compound Q:
  • the C-20 steroid compound of Formula I is Compound R:
  • Compounds A-R exists as a single stereoisomer, wherein the stereochemistry at any center for which stereochemistry is not specified and can be either R or S.
  • the C-20 steroid compounds of Formulas I-IV are believed to be useful for treating, minimizing and/or preventing neuronal damage, such as neuronal damage, resulting from various injuries involving the brain, such as traumatic brain injury (TBI), whether the TBI is mild including concussions, moderate or severe traumatic brain injury.
  • TBI traumatic brain injury
  • the C-20 steroid compounds of Formulas I-IV are believed to be useful to treat and/or prevent MTBI. In another embodiment, the C-20 steroid compounds of Formulas I-IV are believed to be useful to treat and/or prevent concussions.
  • the C-20 steroid compounds of formulas I-IV, especially ent-19 norprogesterone may be administered in a dosage range of from about 0.05 mg/kg to 16 mg/kg, preferably from about 0.05 mg/kg to about 4 mg/kg and even more preferably from about 0.16 mg/kg to about 0.65 mg/kg or from about 1.13 mg/kg to about 45.2 mg/kg per 70 kg patient to treat, minimize and/or prevent TBI, including severe TBI, moderate TBI, mild TBI and concussions, preferably mild TBI, and even more preferable concussions.
  • the C-20 steroid compounds of Formulas I-IV of the present invention can be administered through a number of routes of administration that include, e.g., oral, sublingual, intravenous, intraperitoneal, subcutaneous, intramuscular, intraabdominal, ocular, otic, intranasal, topical, transdermal, subcutaneous and rectal routes of administration.
  • the C-20 steroid compounds can be formulated into, e.g., compositions or admixtures and administered in a dosage form selected from, e.g., a tablet, capsule, gelcap, caplet, powder, granule, liquid, solution, suspension, dispersion, pellet, bead, eyedrop, gel, cream, ointment, salve, balm, lotion or suppository.
  • a dosage form selected from, e.g., a tablet, capsule, gelcap, caplet, powder, granule, liquid, solution, suspension, dispersion, pellet, bead, eyedrop, gel, cream, ointment, salve, balm, lotion or suppository.
  • the present invention contemplates that the C-20 steroid compounds may be administered as a formulation that is swallowed, injected, infused, inhaled, applied transdermally or topically, such as applied to the skin, eyes, ears, nose, lungs, mucosal membranes or any other membrane, or inserted into the rectum. Nonetheless, it should be understood by those versed in the art that preferred routes of administration to treat and/or prevent TBI, especially, mild TBI and concussions, as contemplated by the present invention, is the topical, e.g., pernasal or inhalation, or injection route of administration.
  • the present invention provides a C-20 steroid compounds of Formulas I-IV that is administered through a route selected from oral, sublingual, intravenous, intraperitoneal, ocular, intranasal, transdermal, subcutaneous, and rectal.
  • the C-20 steroid compounds of Formulas I-IV are administered orally.
  • the C-20 steroid compounds of Formulas I-IV are administered sublingually.
  • the C-20 steroid compounds of Formulas I-IV are administered by injection such as intravenously,
  • the C-20 steroid compounds of Formulas I-IV are administered ocularly or otically. In another embodiment, the C-20 steroid compounds of Formulas I-IV are administered intranasally. In another embodiment, the C-20 steroid compound of Formulas I-IV are administered transdermally. In another embodiment, the C-20 steroid compounds of Formulas I-IV are administered subcutaneously. In another embodiment, the C-20 steroid compounds of Formulas I-IV is administered rectally. In another embodiment, the C-20 steroid compounds of Formulas I-IV are administered topically, including by inhalation.
  • the C-20 steroid compounds of Formulas I-IV are administered in a formulation selected from a tablet, capsule, gelcap, caplet, powder, solution, suspension, eyedrop, cream, ointment, lotion, gel or suppository.
  • formulations that contain active agents of Formulas I-IV may optionally contain co-therapeutic agents and inactive excipients.
  • liquid formulations contain a solvent and that said solvent may be either aqueous or organic.
  • the C-20 steroid compounds of Formulas I-IV are administered as a formulation that is swallowed, injected, infused, inhaled, applied topically such as to the skin, eye, mucosal or other membranes and lungs, or inserted into the rectum.
  • a formulation that is swallowed, injected, infused, inhaled, applied topically such as to the skin, eye, mucosal or other membranes and lungs, or inserted into the rectum.
  • solution formulations may be injected, infused, deposited intraperitoneally, deposited subcutaneously, applied to the eye, sprayed or applied into the nose or inhaled as a nebulized liquid or suspension.
  • tablets, capsules, gelcaps and caplets are intended to be swallowed.
  • suppositories are intended for insertion into the rectum while creams, ointments and lotions are intended for topical applications.
  • Scheme 1 represents the formation of compound (9) via two alternative processes.
  • (1) is reacted with (2) to produce (3).
  • the preparation of compound (2) is described in Yamauchi, Noriaki; Natsubori, Yoshiaki; Murae, Tatsushi Bulletin of the Chemical Society of Japan (2000), 73(11), 2513-2519).
  • (3) is subjected to a stereoselective ring closing to form (4).
  • (4) can be converted to (9) either: by selective protection of the carbonyl group to form (5) (as described in Bosch, M.P.; Camps, F.; Coll, J.; Guerrero, T.; Tatsuoka, T.; Meinwald, J. J. Org. Chem.
  • Scheme 2 represents an alternative to the formation of compound (9) of Scheme 1 from the combination of (1) and but-3-en-2-one (43).
  • (1) and (43) are reacted to form (44) which is subjected to a stereoselective ring closing reaction to form (45).
  • (45) is then selectively protected to form (46) (Bosch, M.P.; Camps, F.; Coll, J.; Guerrero, T.; Tatsuoka, T.; Meinwald, J. J. Org. Chem. 1986, 51, 773) which is subjected to a Baylis-Hillman reaction to form (47)
  • the conversion of (47a) to (9), and similar reactions may utilize A1203 as a reagent.
  • activation of a beta-hydroxyketone and subsequent elimination reactions such as those described in Scheme 2 may be accomplished under a variety of conditions including, but not limited to KOH, methanesulfonyl chloride with diisopropylethylamine, para-toluenesulfonyl chloride with dimethylaminopyridine, DCC, pyridinium hydrochloride, alumina.
  • Scheme 3 represents a one step process to form compound (10) by reaction of substituted 2-ethyl-2-methyl-1,3-dioxolane a with ethyl 3-oxobutanoate.
  • leaving group R is -OTs, -OMs, -OTf, -CI, -Br, or -I.
  • leaving group R is -OTs, -Br, or -I.
  • leaving group R is - Br.
  • Scheme 4 represents the formation of compound (14) from the combination of (9) and (10).
  • (9) and (10) are reacted to form (11) which is hydrogenated to form (12).
  • (12) is then double deprotected and cyclized to form (13) which is selectively reprotected to form (14) (Tsunoda, T.; Suzuki, M.; Noyori, R. Tetrahedron Lett. 1980, 21, 1357).
  • Scheme 5 represents the formation of ent-19-Norprogesterone from compound (14) of Scheme 4.
  • (14) is reacted with potassium tert-butoxide and ethyl
  • Scheme 6 represents the formation of compound (15) from the tert-butyl 3-hydroxypent-4- enoate (48) via reduction (Batt, Frederic and labore, Fabienne, European Journal of Organic Chemistry, 2011(30), 6039-6055, S6039/1-S6039/46; 2011), formation of a tosylate and protection with a MOM (Methoxymethyl ether) protecting group to form (49). (49) is then reacted with ethyl 3-oxobutanoate (50) in the presence of a base to form (15).
  • Scheme 7 represents the formation of compound (15) from the tert-butyl 3-hydroxypent-4- enoate (48) via reduction (Batt, Frederic and frae, Fabienne, European Journal of Organic Chemistry, 2011(30), 6039-6055, S6039/1-S6039/46; 2011), formation of a tosylate and protection with a MOM (Methoxymethyl ether) protecting group to form (49). (49) is
  • Scheme 7 represents the formation of ent-19-Norprogesterone from the combination of (9) from Scheme 1 and (15) from Scheme 6.
  • (9) and (15) are reacted in a Robinson annulation to form (16) which is subjected to a Birch reduction or selective hydrogenation reaction to form (17).
  • the MOM ether and ketal of (17) are simultaneously removed to form (18) which is then subjected to a double Wittig reaction to form (19).
  • (19) then undergoes a ring closing metasthesis reaction to form (20) which is subjected to hydroboration reaction to form (21).
  • Double oxidation of (21) results in formation of ent-19-Norprogesterone.
  • Scheme 8 represents the formation of ent-19-Norprogesterone from the combination of (1) from Scheme 1 with a methoxymethylether protected compound (23).
  • (1) and (23) are reacted to form (24) which is subjected to a stereoselective cyclization reaction to form (25).
  • (25) is then selectively protected to form (26) (Tsunoda, T.; Suzuki, M.; Noyori, R. Tetrahedron Lett. 1980, 21, 1357) which is subjected to a Wittig reaction with ethyl triphenylphosphonium bromide to form (27).
  • the MOM ether and the ketal of (27) are simultaneously hydrolyzed to form (28) which is then subjected to a Lewis acid facilitated reduction to form the exocyclic double bond in (29) (Das, Biswanath; Banerjee, Joydeep; Chowdhury, Nikhil; Majhi, Anjoy; Holla, Harish, Synlett (2006), (12), 1879-1882).
  • (29) is subjected to a Robinson annulation with (10) from Scheme 3 to form (30) which is subjected to a Birch reduction or selective hydrogenation to form (31).
  • (31) undergoes a hydroboration reaction to form (32).
  • Hydrolysis of the ketal of (32) with tandem aldol cyclization forms (33). Oxidation of (33) results in ent-19-Norprogesterone.
  • the Lewis acid facilitated reduction is replaced by a
  • Scheme 9 represents an alternative to formation of ent-19-Norprogesterone from Scheme 8.
  • compound (25) is prepared as described in Scheme 8.
  • compound (25) is selectively protected to produce the acetal compound (34) (Tsunoda, T.; Suzuki, M.; Noyori, R. Tetrahedron Lett. 1980, 21, 1357) which is stereoselectively reduced to form the hydroxyl compound (35).
  • (35) is brominated with inversion of stereochemistry to form (36) which is subjected to a nucleophilic displacement with a vinyl anion and inversion of
  • the Lewis acid facilitated reduction is replaced by a
  • Scheme 10 represents the preparation of compound (23) illustrated in Scheme 9. This chemistry is adapted from a protocol for the preparation of a related compound (Batt, F.; horre, F. Eur. J. Org. Chem. 2011, 6039). As illustrated, compound (48) is reduced to compound (50) (Scheme 6). The primary hydroxyl group of compound (51) (Batt, F.; horre, F. Eur. J. Org. Chem. 2011, 6039) is then selectively converted to the corresponding methoxymethyl ether (52). Compound (52) is then oxidized to form compound (23).
  • Scheme 10a represents an alternative to the preparation of compound (23) illustrated in Scheme 10.
  • This chemistry is adapted from a protocol for the preparation of a related compound (Batt, F.; horre, F. Eur. J. Org. Chem. 2011, 6039).
  • propylene glycol is converted to its mono-methoxymethyl ether compound (55).
  • the free hydroxyl group is then oxidized to form the aldehyde of compound (56).
  • the aldehyde is then converted to the allylic alcohol compound (57).
  • Compound (57) is then oxidized to form compound (23).
  • Scheme 11 represents the preparation of compound (2) illustrated in Scheme 1. This chemistry is adapted from a protocol for the preparation of a related compound (Batt, F.; horre, F. Eur. J. Org. Chem. 2011, 6039) and represents an alternative to the synthesis described in Yamauchi, Noriaki; Natsubori, Yoshiaki; Murae, Tatsushi Bulletin of the Chemical Society of Japan (2000), 73(11), 2513-2519). As illustrated, the primary hydroxyl group of compound (51) (Batt, F.; horre, F. Eur. J. Org. Chem. 2011, 6039) is selectively converted to the
  • Scheme 11 a represents an alternative to the preparation of compound (2) illustrated in Scheme 11.
  • This chemistry is adapted from a protocol for the preparation of a related compound (Batt, F.; horre, F. Eur. J. Org. Chem. 2011, 6039) and represents an alternative to the synthesis described in Yamauchi, Noriaki; Natsubori, Yoshiaki; Murae, Tatsushi Bulletin of the Chemical Society of Japan (2000), 73(11), 2513-2519).
  • propylene glycol is converted to its mono-benzyl ether compound (59).
  • the free hydroxyl group is then oxidized to form the aldehyde of compound (60).
  • the aldehyde is then converted to the allylic alcohol compound (61).
  • Compound (61) is then oxidized to form compound (2).
  • Scheme 12 provides an alternative synthesis of Compound (14) as described in Scheme 4.
  • the synthesis includes the sequence converting compound (62) to compound (65) and the conversion of ent-19-nortestosterone (compound 67) to the dioxolane ketal compound (68).
  • (45) is reduced and protected to form (62).
  • (62) is subject to a Baylis- Hillman reaction to form (63) which is further reduced to form (64).
  • (64) is subject to an elimination reaction to form the double bond in (65).
  • (65) is reacted with Compound (10) from Scheme 3 to form (66) which is hydrogenated and cyclized to form ent-19-nortestosterone (67).
  • ent-19-nortestosterone (67) is then ketal protected and reduced to form (14).
  • the conversion of compound (63) to compound (65) is accomplished in a single step comprising a Lewis acid facilitated reduction.
  • Scheme 12a provides an alternative conversion of compound (62) to compound (65).
  • compound (62) is treated with methyl magnesium carbonate (MMC) forming the carboxylated product compound (63a).
  • MMC methyl magnesium carbonate
  • Catalytic hydrogenation reduces the olefin of compound (63a) forming compound (64a).
  • Final decarboxylation in the presence of formaldehyde forms compound (65).
  • the conversion of compound (63a) to compound (64a) and the conversion of compound (64a) to compound (65) are distinct and separate synthetic steps.
  • the conversion of compound (63a) to compound (64a) and the conversion of compound (64a) to compound (65) are run in tandem.
  • catalysts useful for the reduction of a double bond to a single bond including, but not limited to, palladium on carbon, platinum on carbon, palladium hydroxide on carbon, palladium, platinum and Raney nickel.
  • Scheme 13 represents an alternative continuation from compound (13) (Scheme 4) and depends upon the conversion of (13) to the ethyl enol ether compound (70) followed by the Wittig reaction generating compound (71). Reactions of this type are generally described by Antimo, et al., [Steroids 77 (2012) 250-254]. This sequence is completed by initial borane oxidation of (71) followed by hydrolysis of the enol ether and oxidation to form (72). Alternatively, (71) is initially hydrolyzed followed by borane oxidation giving compound (73).
  • Scheme 14 represents an alternative to Scheme 13 and utilizes a reductive silylation to protect the enone of (13) to form (74). Protection of this type is generally described in Iwao, et al. [Tetrahedron Letters 49 (1972) 5085-5038] and Horiguchi, et al. [Journal of the American Chemical Society 111(16) (1989) 6259-6265]. Following borane oxidation of (75) to (77), oxidation of the alcohol and oxidative deprotection of the enone generates ent-19- Norprogesterone. Deprotection of this type is generally described by Yoshihiko, et al. [Journal of Organic Chemistry 43(5) (1978) 1011-1013].
  • silyl enol ether (75) is initially oxidatively converted to (76) followed by borane oxidation to compound (73).
  • the particular process described in the methods of the invention can be utilized to prepare a number of useful intermediates.
  • the intermediates have activity separate and apart from their usefulness in the preparation of ent-Progesterone.
  • the active intermediate compounds have activity in the treatment of traumatic brain injury.
  • the present invention in certain aspects, provides a method for the treatment of traumatic brain injury comprising administering a therapeutically effective amount of an active intermediate compound to a patient in need thereof.
  • active intermediate com ounds include but are not limited to,
  • intermediate B-3 may be represented as Examples
  • the percentage yields reported in the following examples are based on the starting component that was used in the lowest molar amount. Air and moisture sensitive liquids and solutions are transferred via syringe or cannula, and are introduced into reaction vessels through rubber septa. Commercial grade reagents and solvents are used without further purification.
  • the term“concentrated under reduced pressure” refers to use of a Buchi rotary evaporator or equivalent equipment at approximately 15 mm of Hg. All temperatures are reported uncorrected in degrees Celsius (°C). Thin layer chromatography (TLC) is performed on pre-coated glass- backed silica gel 60 A F ⁇ 254250 pm plates.
  • NMR NMR spectra are acquired for each compound when indicated in the procedures below. NMR spectra obtained were consistent with the structures shown. Routine one-dimensional NMR spectroscopy was performed on a 300 MHz Brucker spectrometer. The samples were dissolved in deuterated solvents. Chemical shifts were recorded on the ppm scale and were referenced to the appropriate solvent signals, such as 2.49 ppm for DMSO-d6, 1.93 ppm for CD3CN, 3.30 ppm for CD3OD, 5.32 ppm for CD2Cl2 and 7.26 ppm for CDCI3 for 1H spectra.
  • Equipment used in the execution of the chemistry of this invention include but is not limited to the following:
  • 2-Ethyl-2-methyl-1,3-dioxolane (120mL) and compound 45 (20 g, 1.0 eq.) are combined under nitrogen.
  • Ethylene glycol (1.2 mL, 0.14 eq.) is added followed by p-toluenesulfonic acid (390 mg, 0.02 eq.).
  • the reaction is stirred at 25-30 deg C for 96 hours until the concentration of compound 45 is less than 20% as measured by HPLC.
  • Ethyl acetate (100 mL) is added and the resulting mixture is washed with water (2 x 100 mL), is dried over anhydrous sodium sulfate, is filtered and is concentrated to dryness.
  • Tetrahydrofuran (THF, 10 mL) is added followed by hexamethylphosphoramide (HMPA, 326 mg, 0.25 eq.).
  • HMPA hexamethylphosphoramide
  • Ethyl acetoacetate (1 mL, 1.0 eq.) is added and the mixture is stirred at 0 deg C for 10 minutes.
  • n-Butyllithium 2.5M, 3.6 mL, 1.1 eq.
  • 2-(2-methyl-1,3-dioxolan-2-yl)ethylbromide (1.6 g, 1.0 eq.) is added and the reaction is stirred at 0 deg C for 30 minutes.
  • the reaction is quenched with aqueous oxalic acid (10%, 20 mL) and is washed with dichloromethane (DCM, 3 x 20 mL).
  • the organic phase is additionally washed with saturated aqueous sodium bicarbonate (30 mL) and brine (30 mL).
  • the organic phase is dried over anhydrous sodium sulfate, is filtered and is concentrated.
  • the residue is purified on silica gel (ethyl acetate/hexane 1/30) giving compound 10 (600 mg) as a yellow oil.
  • Ethyl triphenylphosphonium bromide (2.8 g, 3 equivalents) and potassium tert-butoxide (1.0 g, 3.0 equivalents) are combined in anhydrous tert-butanol (10 mL) under nitrogen.
  • the mixture is heated to 75-80 deg C for 20 minutes after which, compound 14 (1.0 g, 1 equivalent) is added.
  • the reaction is stirred at 75 ⁇ 80 deg C for 3 hours after which, it is cooled to 20-25 deg C and is quenched with brine (20 mL).
  • the resulting mixture is washed with ethyl acetate (3 x 20 mL).
  • the combined organic extracts are dried over anhydrous sodium sulfate, are filtered and are concentrated to dryness.
  • the residue is purified on silica gel (10% ethyl acetate/hexane) giving the desired Wittig product in 90% yield.
  • the Wittig product from part (a) (1.0 g, 1 equivalent) is placed under a nitrogen atmosphere and is dissolved in anhydrous tetrahydrofuran (THF, 100 mL). Borane-THF complex (1 M in THF, 3.0 mL, 1 equivalent) is added and the reaction is stirred at 20-25 deg C for 3 hours. The reaction is then concentrated to dryness and sodium hydroxide solution (10% in water, 50 mL) is added followed by hydrogen peroxide solution (30% in water, 0.5 mL). The resulting mixture is stirred at 20-25 deg C for an additional 1 hour after which, water (100 mL) is added. The mixture is then washed with dichloromethane (2 x 100 mL) and the combined organic extracts are washed with brine (50 mL). Concentration of the organic phase yields the crude alcohol which is used in the following step without purification.
  • THF tetrahydrofuran
  • step (b) The crude product from step (b) (2.0 g, 1 equivalent) is dissolved in acetone (20 mL) and hydrochloric acid (30% in water, 20 mL) is added. The reaction is stirred at 20-25 deg C for 30 minutes after which, it is concentrated to dryness. The residue is dissolved in ethyl acetate (50 mL) and water (30 mL) is added. After stirring vigorously for 5 minutes, the phases are separated and the organic phase is washed with saturated aqueous sodium bicarbonate (2 x 25 mL) and brine (25 mL).
  • the resulting mixture is washed with water (10 mL) after which, it is dried over anhydrous sodium sulfate, is filtered and is concentrated to dryness.
  • the residue is purified on silica gel (ethyl acetate/hexane 1/10) giving the desired primary tosylate (500 mg) as a yellow oil.
  • the resulting primary tosylate (100 mg, 1.0 eq.) is dissolved in DCM (10 mL) under nitrogen. Diisopropylethyl amine (DIEA, 1.2 eq.) is added and the mixture is cooled to 0 deg C.
  • DIEA Diisopropylethyl amine
  • 2-Methyl-1,3-cyclopentanedione (3.0 g, 1.2 eq.) is combined with compound 23 (3.1 g, 1.0 eq.) and acetonitrile (ACN, 30 mL).
  • Triethylamine (TEA, 110 mg, 0.05 eq) is added and the reaction is stirred at 25 deg C for 4 hours.
  • Dichloromethane (DCM, 100 mL) is then added and the mixture is washed with aqueous hydrochloric acid (2 x 30 mL) and saturated aqueous sodium bicarbonate (2 x 30 mL). The organic phase is dried over anhydrous sodium sulfate, is filtered and is concentrated to dryness.
  • pro pionaldehyde Compound 55 (1g, 1.0 eq.) is dissolved in dimethylsulfoxide (10 mL) and 2- lodoxybenzoic acid (IBX, 3.5 g, 1.5 eq.) is added. The reaction is stirred at 20 deg C for 12 hours after which, it is washed with saturated aqueous sodium sulfite (20 mL) and is saturated aqueous sodium bicarbonate (20 mL). The organic phase is dried over anhydrous sodium sulfate, is filtered and is concentrated to dryness. The residue is purified on silica gel (ethyl acetate/hexane 1/20) giving compound 56 (0.3 g, 60% purity) as a yellow oil.
  • silica gel ethyl acetate/hexane 1/20
  • Propylene glycol 500 g is combined with benzyl bromide (100 g, 1.0 eq.) under nitrogen.
  • Sodium hydroxide 28 g, 1.2 eq. is added and the mixture is stirred at 20 deg C for 4 hours.
  • Dichloromethane (1.5 L) is added and the mixture is stirred for 20 minutes after which, it is filtered and the filter cake is washed with dichloromethane (600 mL). The combined filtrates are concentrated to dryness giving the desired crude alcohol (328 g). This crude material is dissolved in dichloromethane (3.28 L) and is cooled to -50 deg C. Borontrifluoride etherate (83.1 mL) and phosphoric acid (36.9 mL) are added and the mixture is stirred at -50 deg C for 30 minutes.
  • Isobutylene (2.3 kg) is then added at -45 deg C.
  • the mixture is stirred at -40 deg C for 1 hour after which, it is allowed to warm to room temperature.
  • the reaction is monitored by LCMS during this period until the content of the alcohol is ⁇ 10%.
  • Aqueous ammonium hydroxide (13%, 2.3 L) is then added with vigorous stirring.
  • the layers are separated and the aqueous phase is washed with dichloromethane (1.6 L).
  • the combined organic phases are washed with saturated aqueous ammonium chloride (1.6 L) and brine (1.6 L).
  • the organic phase is dried over anhydrous sodium sulfate, is filtered and is concentrated to dryness.
  • the resulting solids are collected by filtration and are washed with water (50 mL).
  • the solids are collected and are slurried in petroleum ether (150 mL) for 3 hours.
  • the solids are collected by filtration and are washed with petroleum ether (50 mL).
  • the resulting solids are dried in a vacuum oven at 30 deg C for 5 hours yielding compound 63a (30.2 g, 50.4% yield) as a light yellow solid.
  • EXAMPLE 26 Intraperitoneal administration of PRV-002 attenuates motor and cognitive deficits in a rat model of traumatic brain injury
  • the goal of this study is to evaluate the motor and cognitive function of rats treated with PRV-002, an analogue of the enantiomer of progesterone, following traumatic brain injury.
  • Rats receive intraperitoneal injections of either vehicle solution (45% cyclodextrin), PRV-0024mg/kg, or PRV-00216mg/kg at 15 min., 6h, and 24h post-injury.
  • a sham group which does not undergo impact or treatment is used as a control.
  • Motor function is evaluated using a neurobehavioral battery, known as neuroscore, at 24h and 48h post-injury.
  • Cognitive function is assessed using the Morris water maze (MWM) - memory score at 48h post- injury. Time spent swimming in close proximity to the wall of the Morris water maze
  • thigmotaxia is used to evaluate spatial acquisition deficits and potential TBI-induced anxiety.
  • Rats Male Sprague-Dawley rats (Charles River, Wilmington, MA), six weeks of age and weighing between 225— 275 g at the time of injury, are used. Rats are housed in standard Plexiglas cages and are maintained on a 12-12 light cycle with lights on at 0700. Food and water are available ad libitum.
  • rats Prior to surgery, rats are anesthetized via inhalation with an initial induction of 5% isofluorane.
  • the rat’s scalp is shaved and cleaned with a 70% isopropanol solution and 10% betadine solution.
  • anesthesia is maintained at 2.5% isofluorane with oxygen at a rate of 500— 1000 mL/min.
  • the rat’s head is secured in a stereotaxic apparatus and a medial incision is made and the scalp is pulled back with bulldog clips over the frontal bone.
  • a 6 mm circular piece of skull is removed with a Micromotor drill that utilized a removable 6 mm circular drill bit.
  • An electrically-controlled injury device with a 3 mm metal impactor is used to produce the traumatic brain injury.
  • a piston is placed on the dura. Electrical signals from the piston to a transducer signal correct placement.
  • the piston is then used to produce a contusion at a depth of 3 mm. This procedure is used extensively by researchers conducting work on traumatic brain injury and represents one of the most consistent and reproducible forms of injury. Following injury the tissue is closed with 4 ⁇ 0 monofilament sutures. Rats are placed in a heated recovery cage following surgery and are returned to their home cage following recovery.
  • Rats are randomly placed in one of four treatment groups: 1) sham injury group (SHAM), 2) vehicle-treated injury group (VEHICLE), 3) PRV-0024mg/kg-treated injury group (PRV-002 4mg/kg), or 4) PRV-00216mg/kg-treated injury group. Rats receive intraperitoneal injections of either vehicle solution (45% cyclodextrin in sterile water) or PRV-002 solution (PRV-002 powder is dissolved into 45% cyclodextrin solution) at 15 minutes, 6 hours, and 24 hours post-injury. Neuroscore
  • Rats are exposed to a series of four neurobehavioral tests and are observed for abnormal twisting behavior. Rats receive scores from +4 uninjured to (- ) nonfunctional for both left and right forelimbs in the forelimb extension task and forelimb paw placement, the left and right hind limbs in hind limb flexion, and left and right sides for the lateral pulsion test. If no twisting is observed the rat would score as normal +1, and if there is twisting present the rat would score as abnormal (-). The total possible score is 33.
  • the testing criteria is as follows: Forelimb extension
  • Rat extends both forelimbs fully and equally towards floor o +3 Slightly impaired: There is a slight forelimb flexion
  • Rat should resist equally when pushed to each side.
  • Rat does not resist when pushed and falls to its side Forelimb Paw Placement
  • Rat can strongly grasp the object with both paws o +3 Slightly impaired: Rat weakly grasps the object with paw misplacement o +2 Moderately impaired: Rat is weak and unable to maintain grasp of the object
  • Rat is unable to grasp the object
  • Rat shows no attempt to grasp the object Hind limb Flexion
  • Rats have normal extension of hind limbs, no crossing or splaying o +3 Slightly impaired: hind limbs have slight deviation from normal extension, slight clasping or splaying of hind limbs
  • Moderately impaired Moderate crossing over or splaying of hind limbs o +2
  • Modeverely impaired Severe deviation from normal extension with severe crossing over or splaying of hind limbs
  • Hind limbs are crossed or splayed with no normal extension or function Twisting
  • Thigmotaxis is a measure of the amount of time rats spend“wall hugging” or swimming around the edge of the tank. Time spent traveling in the thigmotaxia area is measured and is indicative of high anxiety and spatial acquisition deficits in injured animals. See FIG.2. Statistical Analysis
  • a one-way analysis of variance is used to evaluate group differences in MWM memory score and MWM thigmotaxia.
  • post-hoc analysis of pair-wise comparisons is carried out using Fisher’s Protected Least Significant Differences (PLSD) test.
  • Neuroscore data is analyzed using the Kruskall-Wallis test to evaluate group differences.
  • pair-wise comparisons are carried out using the Mann-Whitney U Test. Results
  • a one-way analysis of variance is used to evaluate group differences in MWM memory score.
  • Post-hoc analysis shows that vehicle-treated injured rats have significantly worse cognitive function than sham, PRV-0024mg/kg-, and PRV-00216mg/kg- treated rats during both trials of the MWM task. See FIG.5A and FIG.5B.Morris Water Maze — Time Spent in Thigmotaxia. See FIG.2.
  • a one-way analysis of variance is used to evaluate group differences in time spent in thigmotaxia during the MWM task.
  • Post-hoc analysis shows that vehicle-treated injured rats spend significantly more time in thigmotaxia than sham, PRV-002 4mg/kg-, and PRV-00216mg/kg-treated rats during both trials of the MWM task. See FIG.6A and FIG.6B. Discussion
  • Neuroscore, MWM-memory score, and MWM-time spend in thigmotaxia all reveal significant motor and cognitive deficits in vehicle-treated rats following experimental traumatic brain injury. Though no significant group differences are seen in neuroscore at 24h post-injury, by 48h rats treated with PRV-0024mg/kg or PRV-00216mg/kg show significant attenuation of TBI-induced motor function deficits. Rats treated with PRV-0024mg/kg or PRV-00216mg/kg show amelioration of TBI-induced cognitive deficits, as measured by the MWM-memory score at 48h hours post-injury.
  • the decreased time spent in thigmotaxia may also indicate that treatment with PRV-0024mg/kg or PRV-00216mg/kg may induce anxiolytic effects following TBI.
  • Example 27 Intranasal administration of PRV-002 attenuates motor and cognitive deficits in a rat model of traumatic brain injury
  • the goal of this study is to evaluate the motor and cognitive function of rats treated via intranasal administration with PRV-002, an analogue of the enantiomer of progesterone, following traumatic brain injury.
  • an anatomical evaluation is performed using PRV002 labeled with Evans Blue dye to determine the optimal intranasal/intracerebral penetration of compound using intranasal administration via a miniature atomizer vs. a manual pipette.
  • Post-mortem evaluation determined a clear advantage of the miniature atomizer over the pipette technique with respect to maximal nasal mucosal penetration.
  • a sham group which did not undergo impact or receive treatment was used as a control.
  • Motor function is evaluated using a neurobehavioral battery, known as neuroscore, at 24h and 48h post-injury.
  • Cognitive function is assessed using the Morris water maze (MWM) - memory score at 48h post-injury.
  • MFM Morris water maze
  • thigmotaxia Time spent swimming in close proximity to the wall of the Morris water maze (thigmotaxia) is used to evaluate spatial acquisition deficits and potential TBI- induced anxiety.
  • Rats Male Sprague-Dawley rats (Charles River, Wilmington, MA), six weeks of age and weighing between 225— 275 g at the time of injury, are used. Rats are housed in standard Plexiglas cages and are maintained on a 12-12 light cycle with lights on at 0700. Food and water are available ad libitum.
  • rats Prior to surgery, rats are anesthetized via inhalation with an initial induction of 5% isofluorane.
  • the rat’s scalp is shaved and cleaned with a 70% isopropanol solution and 10% betadine solution.
  • anesthesia is maintained at 2.5% isofluorane with oxygen at a rate of 500—1000 mL/min.
  • the rat’s head is secured in a stereotaxic apparatus and a medial incision is made and the scalp is pulled back with bulldog clips over the frontal bone.
  • a 6 mm circular piece of skull is removed with a Micromotor drill that utilized a removable 6 mm circular drill bit.
  • An electrically-controlled injury device with a 3 mm metal impactor is used to produce the traumatic brain injury.
  • a piston is placed on the dura. Electrical signals from the piston to a transducer signal correct placement.
  • the piston is then used to produce a contusion at a depth of 3 mm. This procedure is used extensively by researchers conducting work on traumatic brain injury and represents one of the most consistent and reproducible forms of injury. Following injury the tissue is closed with 4 ⁇ 0 monofilament sutures. Rats are placed in a heated recovery cage following surgery and are returned to their home cage following recovery.
  • mice receive an IN spray of either vehicle solution (45% cyclodextrin in sterile water) or PRV- 002 solution (PRV-002 powder dissolved into 45% cyclodextrin solution) at 15 minutes, 6 hours, and 24 hours post-injury using a micro atomizer.
  • vehicle solution 45% cyclodextrin in sterile water
  • PRV- 002 solution PRV-002 powder dissolved into 45% cyclodextrin solution
  • Rats are exposed to a series of four neurobehavioral tests and are observed for abnormal twisting behavior. Rats receive scores from +4 uninjured to (- ) nonfunctional for both left and right forelimbs in the forelimb extension task and forelimb paw placement, the left and right hind limbs in hind limb flexion, and left and right sides for the lateral pulsion test. If no twisting is observed the rat would score as normal +1, and if there is twisting present the rat scores as abnormal (-). The total possible score is 33.
  • the testing criteria is as follows: Forelimb extension Suspend the rat by its tail and determine the forelimb extension toward floor. Score separately for both the left and right forelimb.
  • Rat extends both forelimbs fully and equally towards floor o +3 Slightly impaired: There is a slight forelimb flexion
  • Rat should resist equally when pushed to each side.
  • o +3 Slightly impaired Rat maintains moderate resistance
  • Rat does not resist when pushed and falls to its side
  • Rat can strongly grasp the object with both paws o 0 +3 Slightly impaired: Rat weakly grasps the object with paw misplacement o 0 +2 Moderately impaired: Rat is weak and unable to maintain’ grasp of the object
  • Rat is unable to grasp the object
  • Rats have normal extension of hind limbs, no crossing or
  • Moderately impaired Moderate crossing over or splaying of hind limbs o 0 +1
  • Severely impaired Severe deviation from normal extension with severe crossing over or splaying of hind limbs
  • Hind limbs are crossed or splayed with no normal extension or function Twisting
  • a one-way analysis of variance is used to evaluate group differences in MWM memory score and MWM thigmotaxia.
  • post-hoc analysis of pair-wise comparisons is carried out using Fisher’s Protected Least Significant Differences (PLSD) test.
  • Neuroscore data is analyzed using the Kruskall-Wallis test to evaluate group differences.
  • pair-wise comparisons are carried out using the Mann-Whitney U Test. Results
  • a one-way analysis of variance is used to evaluate group differences in MWM memory score.
  • Post-hoc analysis shows all groups have significantly lower memory scores than PRV-0024mg/kg-treated rats during trial 1. See FIG.8A and FIG.8B.
  • a one-way analysis of variance is used to evaluate group differences in time spent in thigmotaxia during the MWM task.
  • Post-hoc analysis shows that sham and PRV-0024mg/kg-treated rats spent significantly less time in thigmotaxia, compared to vehicle-treated rats. See FIG.9A and FIG.9B.
  • PRV-0020.1 mg/kg, or PRV-0024mg/kg have significantly better motor function, as compared to vehicle-treated rats at 48h post-injury.
  • PRV-0020.05mg/kg- and PRV-0021 mg/kg- treated rats have significantly worse performance, compared to sham rats at 48h post-injury (Table 4). See also FIG.10 and FIG.11. Table 3. Neuroscore— 24h Post-Injury
  • Neuroscore, MWM-memory score, and MWM-time spent in thigmotaxia all reveal significant motor and cognitive deficits in brain-injured, vehicle-treated rats following
  • Post-traumatic motor deficits at 24 h post-injury are significantly improved in animals treated with either PRV002 (0.1mg/kg) or PRV002 (4mg/kg).
  • PRV002 0.1mg/kg
  • PRV002 0.1mg/kg
  • PRV002 4mg/kg
  • the results of this study reveal the efficacy of PRV-002 in counteracting TBI-induced motor and cognitive deficits in the cortical impact model of TBI in rats.
  • Apparent solubility of PRV-002 in 35.8% of 2- Hydroxypropyl -6-cyclodextrin is about 23 mg/ml.
  • the PRV-002 solution is prepared by adding about 30 mg of PRV-001 compound to about 1 ml of 35.8% 2-Hydroxypropyl -p-cyclodextrin. Solution is hazy after mixing for about 60 min. Then about 0.1 ml of about 35.8% 2-Hydroxypropyl -P-cyclodextrin is added to about 1 ml of PRV-002—Cyclodextrin mixture. Solution is hazy after mixing for about 120 min. Additional 0.1 ml of about 35.8% 2-Hydroxypropyl -p-cyclodextrin is added to about 1.1 ml of PRV-002— Cyclodextrin mixture and left on mixing overnight (0/N).
  • This project is to perform a polymorph screening for compound PRV- 002. This screening is designed to evaluate polymorphism in the compound and select an appropriate crystalline phase for development with associated risks analyzed.
  • Type A Using anhydrous Type A as the starting material, a polymorph screening is performed under 107 conditions through methods of anti-solvent addition, evaporation, slow cooling, slurry conversion, vapor diffusion, polymer-induced crystallization and grinding. A meta-stable anhydrate, Type B, and amorphous phase are identified, both of which will convert into Type A at ambient conditions.
  • Type A is an anhydrous form suitable for further development. Crystal morphology control is recommended as formation of large rods, observed from some screening conditions, may cause decreased flowability in down-stream processes. Characterization of solid forms
  • Type B amorphous convert back to Type A.
  • a brief inter-conversion diagram is illustrated in FIG.13.
  • Type A is recommended as the stable anhydrous form for further development. See Figs. 12 and 13.
  • PRV-002 Type A ent-19-norprogesterone
  • the starting material (Batch PH-PRV-1302-GLP-0B-A-1, with a Crystal Pharmatech ID of 807302- 25-A) as-received is crystalline, with the crystal form named as Type A.
  • X-ray powder diffraction (XRPD) pattern is displayed in FIG. 14.
  • TGA Thermogravimetric analysis
  • DSC differential scanning calorimeter
  • PLM Polarized light microscope
  • Type B (ent-19-norprogesterone)
  • Type B can be generated via slow evaporation in IPAc or THF/n-heptane (about 4:1, v/v) solution at about RT.
  • Type B sample (807302-42-A) is obtained via well-controlled evaporation of THF/n-heptane (about 4:1, v/v) solution at about RT. Solids are isolated and characterized before the solution is evaporated to dryness. XRPD pattern is shown in FIG. 19 and
  • TGA/DSC data are displayed in FIG. 20.
  • a weight loss of about 1.4% up to about 150 oC is observed in TGA, and one exotherm at about 51.2 oC is observed before melting at about 142.3 oC (onset temperature) in DSC.
  • the XRPD pattern collected after sample 807302-42-A is stored at ambient conditions for about 5 hours conformed to Type A. This indicates Type B is meta-stable to Type A and converts readily at ambient conditions. The exothermic event in the DSC of Type B is believed to be most likely caused by the phase transition from Type B to Type A.
  • Precipitates (807302-27-A8) appear after adding H2O into DMSO solution, and XRPD pattern in FIG . 22 reveals that amorphous sample is obtained. Characteristic peaks of Type A can be observed from FIG.23, after sample 807302-27-A8 is kept at ambient conditions for about two weeks.
  • a total of 20 anti-solvent addition experiments are carried out. About 15 mg of Type A sample (807302-25-A) is dissolved in about 0.2-2.2 mL solvent to obtain a clear solution, and the solution is magnetically stirred, then followed by addition of the relative anti-solvent to induce precipitation or the total amount of anti-solvent reaches about 15.0 mL. The precipitate is isolated for XRPD analysis. Clear solutions are transferred to agitation at about 5 oC for about 4 days, and solids are then tested by XRPD. The final clear solutions are transferred to evaporation at about RT. XRPD patterns are displayed from F I G . 24. F I G . 25 , F I G . 26 a n d F I G . 27. Results are summarized in Table 8, which show that only Type A is produced. Table 8. Summary of anti-solvent addition experiments Reverse anti-solvent addition experiments Reverse anti-solvent addition
  • Type A sample 807302-25-A
  • a saturated solution is added into a 20-mL glass vial with about 6 mL of relative anti-solvent and mixture is stirred at about RT to induce precipitation.
  • the precipitate is agitated for about 30 seconds and isolated for XRPD analysis.
  • the final clear solutions are transferred to evaporation at about RT.
  • XRPD patterns are displayed in FIG.28 and FIG.29. Results are summarized in Table 9, which show that no new crystal form is produced.
  • Type A sample 807302-25-A
  • solvent solvent in a 1.5-mL glass vial.
  • the remaining solids are isolated for XRPD analysis. XRPD patterns are displayed in FIG. 35 and FIG.
  • Type A sample 807302-25-A
  • a 3-mL vial which is placed into a 20-mL vial with about 2 mL of volatile solvent.
  • the 20-mL vial is sealed with a cap and kept at about RT for about 2 days allowing solvent vapor to interact with sample.
  • XRPD patterns are displayed in FIG. 37 and F I G . 38.
  • the solids are tested by XRPD and the results are summarized in Table 13, which indicate no form change.
  • Polymer-induced crystallization experiments are performed with two sets of polymer mixtures in five different solvents. Approximate 15 mg of Type A sample (807302-25-A) is dissolved in about 1.0 mL of appropriate solvent to obtain a clear solution in a 3-mL vial. About 2 mg of polymer mixture is added into 3-mL glass vial. All the samples are sealed using parafilm and then are transferred to evaporation at about RT to induce precipitation. The solids are isolated for XRPD analysis. XRPD patterns are displayed in F I G . 40 and F I G . 41.
  • Type A sample 807302-25-A is dissolved in about
  • FIG. 42 and FI G. 43. Results are summarized in Table 16, which indicate
  • Type A sample 807302-25-A
  • the solid is analyzed by XRPD and, as shown in FIG. 44, only Type A is obtained.
  • the objective of this study is to evaluate the toxicity and toxicokinetics of PRV-002, when administered intranasally at doses three times a day, approximately 4 hours apart, for 14 days at concentrations of about 0 mg/mL, about 3 mg/mL, about 10 mg/mL or about 23 mg/mL at a volume of about 1 mL/nostril. 1 See Table 18. Reversibility of toxicity is evaluated during a 14-day recovery period following the final dose of test article, and systemic exposure is evaluated. Observations and conclusions
  • PRV-002 does not affect ophthalmology, body weights or food consumption.
  • Dose Preparation Weekly (if supported by stability data). Dose concentration is confirmed by analytical methods on Days 1, 14.
  • Body Weight Measured prior to the study, weekly thereafter, and just prior to
  • Ophthalmic Exams Complete ophthalmic exams will be performed on all available study animals prior to the study and just prior to all scheduled necropsies. Electrocardiograms: Standard 6-lead ECG to be performed on all available animals prior to the study and post dose on Day 1 and post dose one day prior to study termination.
  • Clinical Pathology Routine panels of serum chemistry, hematology, and coagulation
  • TK Samples Blood samples (about 1.0 mL) will be collected (in sodium heparin;
  • Organ weights Adrenals, brain, heart, kidneys, liver, lungs, ovaries with oviducts,
  • Histopathology All animals. A full tissue list (approximately 66 tissue sections) and gross lesions from all animals
  • the objective of this study is to evaluate the toxicity of PRV-002, when it is administered as three times a day doses, approximately 4 hours apart, for 14 days at concentrations of about 0 mg/mL, about 3 mg/mL, about 10 mg/mL or about 23 mg/mL at a volume of about 50 ⁇ L/nostril. Reversibility of toxicity is evaluated during a 14-day recovery period following the final dose of test article, and systemic exposure is evaluated. Observations and conclusions
  • nose/nares staining/crusty is observed in four, eleven and twenty-two PRV-002 treated animals at concentrations of about 3 mg/mL, about 10 mg/mL, and 23 mg/mL, respectively. See Table 19.
  • the combined finding of nose/nares staining/crusty is considered PRV-002-related, but not adverse, as it did not affect the overall well-being of the animals.
  • Mouth staining is observed at a higher incidence at about 23 mg/mL and is considered PRV-002-related. No PRV-002 animals are remarkable for these findings during the recovery phase.
  • PRV-002 affected body weight in males at about 23 mg/mL.
  • group mean male body weight is lower (about -9%) at about 23 mg/mL, as when compared to the control group mean.
  • group mean body weight gain is lower (about -2.1 grams) at about 23 mg/mL, as when compared to the control group mean (about +26.4 grams). These differences are not considered adverse.
  • Recovery group mean values in the about 23 mg/mL male group are generally similar to the control group means.
  • Group mean food consumption is slightly lower in males at about 23 mg/mL during the dosing and recovery phases, as when compared to the control group means. This difference is not considered adverse.
  • Body weights prior to dose administration, weekly thereafter and at termination.
  • Hematology Pretrial, prior to first dose and at termination.
  • ®Hydroxypropyl Beta Cyclodextrin solution on the day of treatment.
  • PRV-002 intended doses of about 3 mg/mL, about 10 mg/mL and about 23 mg/mL.
  • the vehicle or test article is dosed on three occasions, approximately 4 hours apart, by administering about 1 mL to each nostril. Animals are observed once daily and on days of dose
  • One-minute means of hemodynamic parameters as well as ECG parameters are measured for a period of at least 22 hours following each dose. The following parameters are analyzed using the D.I.S.S. CA Recorder Systems Version 3.0.1: Body temperature, Systolic Arterial Pressure (SAP), Diastolic Arterial Pressure (DAP), Mean Arterial Pressure (MAP), Heart Rate (HR), P duration, PR Interval, QRS Interval, R amplitude, and QT Interval.
  • SAP Systolic Arterial Pressure
  • DAP Diastolic Arterial Pressure
  • MAP Mean Arterial Pressure
  • HR Heart Rate
  • P duration PR Interval
  • QRS Interval QRS Interval
  • R amplitude QT Interval.
  • a board-certified veterinary cardiologist examined one-minute tracings of the ECGs at about 15 minutes prior to dosing and at about 30 minutes post dose and at about 1, about 2, about 4, about 8, about 12 and about 22 hours post dose.
  • Intranasal administration of PRV-002 at doses of about 3 mg/mL, about 10 mg/mL or about 23 mg/mL does not induce any effects on heart rate or blood pressure in conscious telemetered beagle dogs nor does it have any toxicologic effects on cardiac rhythm or ECG morphology in this study.
  • Bioanalysis of the plasma samples that are collected on each day of dosing at about 10 minutes following the third dose confirms exposure to PRV-002 in a dose-dependent manner.
  • Intranasal administration of PRV-002 three times daily at doses of about 3 mg/mL, about 10 mg/mL or about 23 mg/mL to conscious telemetered dogs does not have any effects on heart rate, arterial blood pressure, cardiac rhythm or ECG morphology.
  • FIGS.45A-45D Certain cardiovascular parameters are depicted in FIGS.45A-45D.
  • FIGS.46A-46D The effects of intranasal administration of about 10 mg/mL PRV-002 upon certain cardiovascular parameters are depicted in FIGS.46A-46D.
  • FIGS.47A-47D The effects of nitranasal administration of about 23 mg/mL PRV-002 upon certain cardiovascular parameters are depicted in FIGS.47A-47D. Table 20– Clinical Observations
  • CSF spinal fluid
  • Numbers represent PRV-002 levels in various parts of the brain in dogs that are dosed IN 3 times at about 4 hour intervals in one day. Dogs are sacrificed and tissues are harvested at about 30 minutes after the last dose. Dogs are given about 1 mL/nostril at each dosing interval for a total dose of about 46 mg/dog per dosing interval. See Table 29. Table 29. Dog Brain Bio-Distribution: Numbers represent brain PTV-002 levels
  • the nano-formulation is a nanosuspension of compound PRV-1302 in about 1% Kolliphor P338/P188.
  • the final PRV-1302 formulation had an API concentration of about 23.4 mg/mL (PRV-1302) with a particle size of approximately 450 nm 2 .
  • Quantitative determination of compound PRV-1302 is accomplished using the HPLC method provided to us. A final average concentration of about 23.4 mg/mL is measured for the PRV-1302 formulated suspension. See Table 30. Table 30. Nanosuspension formulation specifications
  • PRV-002 Concentration (PRV-1302 content by HPLC): about 23.5 mg/mL
  • PRV-002 Particle size of formulated PRV-1302: about 450 nm
  • PRV-002 (Pharmaron: PH-PRV-1302-0B) / PH-PRV-1302-GLP-0B-A-1
  • PRV-002 Concentration: about 23 mg/mL 2
  • the present invention contemplates a nanoparticle size range for PRV-002 (ent-19-norprogesterone) of about 400 nm to about 450 nm.
  • Vehicle 2-hydroxypropyl- ⁇ -cyclodextrin (45% aqueous solution) Dog CSF bio-distribution
  • Numbers represent PRV-002 levels in CSF of dogs that are dosed IN 3 times at about 4 hour intervals in one day. Dogs are anesthetized and CSF is harvested at about 30 minutes after the last dose. Dogs are given about 1mL/nostril at each dosing interval for a total dose of about 46 mg/dog per dosing interval. See Table 32. Table 32. Dog PRV-002 (ent-19-norprogesterone) CSF Bio-Distribution
  • Numbers represent PRV-002 (ent-19-norprogesterone) levels in plasma of dogs that are dosed IN 3 times at 4 hour intervals in one day. Dogs are given about 1mL/nostril at each dosing interval for a total dose of about 46 mg/dog per dosing interval. See Table 33 and FIG.48.
  • PRV-002 ent-19-norprogesterone
  • ICH M3 R2
  • the GLP repeat dose toxicology studies included safety pharmacology elements (CNS in rat, CV/respiratory in dog).
  • a dog brain bio-availability study was conducted in dogs to demonstrate if the drug reaches the brain (the target organ) and if minimal drug amounts circulate systemically when given by IN route.
  • the GLP toxicology program was conducted at BASi in Evansville, IN as follows: A 14- day rat toxicology study was done. The objective of this study was to evaluate the toxicity of PRV-002, when administered three times a day, approximately 4 hours apart, for 14 days at concentrations of 0, 3, 10 or 23 mg/mL at a volume of 50 ⁇ L/nostril. Reversibility of toxicity was evaluated during a 14-day recovery period following the final dose of test article, and systemic exposure was evaluated. During the dosing phase, nose/nares staining/crusty was observed in four, eleven and twenty-two PRV-002 treated animals at concentrations of 3, 10, and 23 mg/mL, respectively. The combined finding of nose/nares staining/crusty was considered PRV-002-related, but not adverse, as it did not affect the overall well-being of the
  • PRV-002 affected body weight in males at 23 mg/mL. On Day 14, group mean male body weight was lower (-9%) at 23 mg/mL, compared to the control group mean. On Days 1-14, group mean body weight gain was lower (-2.1 grams) at 23 mg/mL, compared to the control group mean (+26.4 grams). These differences were not considered adverse.
  • Recovery group mean values in the 23 mg/mL male group were generally similar to the control group means. Group mean food consumption was slightly lower in males at 23 mg/mL during the dosing and recovery phases, compared to the control group means.
  • a 14-day dog toxicology study was conducted. The objective of this study was to evaluate the toxicity of PRV-002, when administered three times a day, approximately 4 hours apart, for 14 days at concentrations of 0, 3, 10 or 23 mg/mL at a volume of 1
  • PRV-002 did not affect ophthalmology, body weights or food consumption. Increased salivation was observed in all combined male and female PRV-002 treated groups, with incidence increased with concentration and is considered PRV-002-related, but not an adverse effect.
  • PRV-002 did not affect ophthalmology, body weights or food consumption. Increased salivation was observed in all combined male and female PRV-002 treated groups, with incidence increased with concentration and is considered PRV-002-related, but not an adverse effect.
  • PRV-002 did not affect ophthalmology, body weights or food consumption. Increased salivation was observed in all combined male and female PRV-002 treated groups, with incidence increased with concentration and is considered PRV-002-related, but not an adverse effect.
  • PRV-002 did not affect ophthalmology, body weights or food consumption. Increased salivation was observed in all combined male and female PRV-002 treated groups, with incidence increased with concentration and is considered PRV-002-related, but not an adverse effect.
  • a dog brain bio-availability study was conducted as follows: A nanoparticle formulation of PRV-002 was tested in dogs to determine if it is absorbed into the target organ (the brain) after intranasal administration. Three dogs each weighing approximately 6 kg were dosed intranasally. The dogs were given 46 mg of drug at 4-hour intervals three times in one day. Blood samples were taken to determine levels of the drug at 10, 30, 60 minutes and 2, 4, and 8 hours post dose. Cerebral spinal fluid (CSF) samples were taken along with the brain for bioanalysis at necropsy approximately 30 minutes after the last dose. The results showed excellent absorption and distribution into all regions of the brain. In addition, measurable amounts of the drug were found in the CSF. Ninety-three (93)% of the recovered drug was found in the brain with only 5% found in the circulating plasma. Approximately 2% of the recovered drug was found in the CSF. See Table 29 above and Table 34.
  • CSF Cerebral spinal fluid
  • PRV-002 ent-19-norprogesterone
  • Numbers represent PRV-002 (ent-19-norprogesterone) plasma levels
  • a dry powder nasal formulation will facilitate ease of administration in the field of play or the field of battle.
  • the capability to administer this product both prophylactically and following injury gives its use and marketing ability a significant advantage.
  • a dry powder formulation is a practical, stable, field- deliverable compound—suitable for application in a sports field, or for military use. Summary Product Profile
  • Neurosteroids reduce inflammation after TBI through CD55 induction. Neurosci Lett,
  • ProTECT A Randomized Clinical Trial of Progesterone for Acute Traumatic Brain Injury. Ann Emerg Med, 49 (4):391-402 (2007) 57. Xiao, G., J. Wei, W. Yan, W. Wang, and Z. Lu, Improved outcomes from the administration of progesterone for patients with acute severe traumatic brain injury: a randomized controlled trial. Crit Care, 12(2):R61 (2008). 58. Sayeed, I., Q. Guo, S.W. Hoffman, and D.G. Stein, Allopregnanolone, a progesterone metabolite, is more effective than progesterone in reducing cortical infarct volume after transient middle cerebral artery occlusion.
  • Facr/020Sheet4Pros-a.pdf 73 Victimization of Persons with Traumatic Brain Injury or Other Disabilities: A Fact Sheet for Friends and Families at http://www.cdc.gov/trau maticbraini njury/pdfNictimization Person Facr/020Sheet4FrdsFa 1-a.pdf 74. Report to Congress on Traumatic Brain Injury in the United States: Understanding the Public Health Problem among Current and Former Military Personnel (June 2013) at

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Abstract

La présente invention concerne des composés stéroïdes C-20, des compositions et des méthodes d'utilisation de ceux-ci pour traiter, atténuer les symptômes et/ou prévenir un traumatisme craniocérébral (TCC), y compris un TCC grave, modéré ou léger.
EP16815158.7A 2015-06-21 2016-06-21 Composés stéroïdes c-20, compositions et utilisation de ceux-ci pour traiter un traumatisme craniocérébral (tcc), y compris des commotions cérébrales Withdrawn EP3416651A1 (fr)

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KR102396328B1 (ko) 2013-04-17 2022-05-10 세이지 테라퓨틱스, 인크. 19-노르 c3,3-이치환된 c21-n-피라졸릴 스테로이드 및 그의 사용 방법
US20160068563A1 (en) 2013-04-17 2016-03-10 Boyd L. Harrison 19-nor neuroactive steroids and methods of use thereof
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US10246482B2 (en) 2014-06-18 2019-04-02 Sage Therapeutics, Inc. Neuroactive steroids, compositions, and uses thereof
TW201625660A (zh) * 2014-09-17 2016-07-16 普瑞弗克斯公司 Ｃ－２０類固醇化合物、其組成物及用於治療包含震盪之創傷性腦損傷（ｔｂｉ）的用途
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